Abstract: Background Spinal cord injury (SCI) due to lack of restoration of damaged axons is associated with sensorimotor impairment. This study was focused on using the human Placental mesenchymal stem cells- exosome (hPMSCs- exosomes) in an animal model of severe SCI under a new myelogram protocol to confirm lumbar puncture (LP) injection accuracy and evaluate intrathecal space. Methods Mesenchymal stem cells (MSC) were extracted from human placenta tissue and were characterized. HPMSCs- exosomes were isolated by ultracentrifuge. After creating the severe SCI model, LP injection of exosomes was performed in the acute phase. Myelogram was also employed. The improved functional recovery of the animals in the treatment and control groups was followed by recording movement scores for 6 weeks. Hematoxylin-Eosin (H&E) staining was used to evaluate to detect pathological changes and glial scar size. The Immunohistochemistry (IHC) of GFAP and NF200 factors as well as the apoptosis tunnel test was investigated in the tissue samples from the injury site Results The results demonstrated that the use of the myelogram can be a feasible, appropriate and cost-effective method to confirm the accuracy of therapeutic agents LP injection and examine the subarachnoid space in the model of laboratory animals. Furthermore, intrathecal injection of hPMSCs-exosomes in the acute phase of SCI can improve motor function by attenuating apoptosis of neurons at the site of injury, decreased GFAP expression and increased NF200 in the treatment group, reducing glial scarring, and increasing axonal regeneration. Improving functional recovery by not creating bedsores in the treatment group and preventing hematuria were other effects of the exosome Conclusions In conclusion, the effects of hPMSCs-exosome can be considered to be not only in restoring function but also in preventing complications and managing symptoms. Thus, the neuroregenerative and anti-apoptotic potential of hPMSCs-exosome can be considered a therapeutic approach in SCI reconstructive medicine.

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Human placental mesenchymal stem cells derived exosomes improved functional recovery via attenuating apoptosis and increasing axonal regeneration...

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Glioblastoma multiforme (GBM) or simply glioblastoma, is a type of cancer characterized by the growth of an aggressive neoplasm (tumor) in the brain or spinal cord. This type of cancer often occurs in older adults, although the younger population may also be affected.

Read Also: Targeting Hox Gene Dysregulation a Promising Approach for the Treatment of Glioblastoma Multiforme

Glioblastoma

This cancer type is known to be difficult to treat because of its high tendency to reoccur in patients, even after the combination of the three known procedures to treat cancers: surgery, radiotherapy, and chemotherapy. Glioblastoma has been a thorn in the flesh in the world of medicine amongst all cancer types due to the low survival rate of patients affected by it (average survival of 18 months, with only 5% of patients living up to five years). The following factors make this possible: no specific signs or symptoms are noticed leading to late diagnosis and the ability of the cancer cells to resist treatment procedures (the major factor).

Studies have been ongoing to uncover the mechanism behind this major factor, and it has been revealed that Glioblastoma multiforme contains a functional subset of cells known as glioblastoma stem-like cells (GSCs) which are the brain behind its reoccurrence capacity. The identity of these cells remained hidden until a recent study done by a group of scientists finally uncovered it.

Read Also: Brain Cancer: A Promising New Treatment for Destroying Aggressive Glioblastoma Cell

The team found out that these functional subsets of cells can be identified through singular mitochondrial alternative metabolisms. After intensively studying the metabolic reactions of these cells, they developed a tumor model that possessed the features of the GBM cultured in the lab. This way, they discovered that GSCs use these two metabolic reactions alpha-ketoglutarate reductive carboxylation and pyruvate carboxylation within their cells. They also discovered that these reactions are catalyzed by the enzymes isocitrate dehydrogenase and pyruvate carboxylase respectively.

They were able to uncover that their high rate of survival which facilitated the recurrence of the tumor is linked to the pyruvate carboxylation reaction. This discovery is important as it means that doctors may now be able to tackle the reoccurring ability of the tumor effectively.

It has always been known that treating glioblastoma is difficult due to its high recurring ability. However, with the revelation from this study, it is now possible for physicians to come up with more effective treatment procedures that would result in a reduced recurrence of the tumors, and an increased survival rate of patients.

This study raises the hopes of both physicians and patients as it reveals a way to hinder the recurrence of glioblastoma tumors. More research is still ongoing to hasten the innovation of a more effective treatment technique.

Read Also: Brain Cancer: Researchers Reprogram Immune Cells to Improve the Effectiveness of Treatment

Pyruvate carboxylation identifies Glioblastoma Stem-like Cells opening new metabolic strategy to prevent tumor recurrence

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How the Regenerative Properties of Glioblastoma Can Be Terminated - Gilmore Health News

Spinal Cord Stem Cells Comments Off on How the Regenerative Properties of Glioblastoma Can Be Terminated – Gilmore Health News | August 2nd, 2022

LAS VEGAS, NV, Aug. 01, 2022 (GLOBE NEWSWIRE) -- via NewMediaWire Meso Numismatics, Inc. (Meso Numismatics or the Company) (MSSV), a technology company specializing in Biotech and Numismatics, is pleased to announce additional global expansion by opening stem cell therapy and regenerative medicine facilities in Indonesia. The new facilities emphasize Global Stem Cells Group's objective of introducing its therapies and technology to meet market demands in populous parts of the world.

In partnership with the Dr. Yanti Aesthetic Clinics, which currently has 6 branches across Indonesia, this latest GSCG expansion will promote high standards of service in regenerative medicine across the country. As part of this effort, through GSCG the International Society for Stem Cells Applications (ISSCA) has granted Dr. Yanti Aesthetic Clinics membership and use of its brand, products, therapies, and training on how to apply stem cell therapies.

This new partnership seeks to expand the Global Stem Cells Group (GSCG) brand and create centers of excellence in cell therapy to meet the high demand within the vast Asian markets, said David Christensen, CEO of MSSV. GSCG is rapidly expanding its global operations as it seeks to become a significant player in the lucrative regenerative medicine industry. To achieve our expansion plans, our organization is partnering with healthcare providers specializing in regenerative medicine with at least five years of experience in the healthcare sector.

Video: https://youtu.be/T2CFjsps9qk

The vision behind the effort.

The Indonesia addition is the latest part of an expanding medical network of partners, and it will formalize and strengthen ties, establishing a global center of excellence to guarantee that we effectively use the underlying basic stem cell technology for medical conditions, where traditional therapeutic approaches seem to have failed. This is consistent with GSCG's overall strategy for developing regenerative medicine through data-driven studies, disease modeling, and cell-based therapeutics.

The Dr. Yanti Aesthetic Clinic is a key partnership because it provides the organizational and physical infrastructure needed to disseminate need-based stem cell locally. And Global Stem Cells Group's outstanding cell and stem cell biology and disease pathophysiology give an edge to patients for which they are prescribed.

The opening in Indonesia also presents the perfect opportunity to translate breakthrough therapies from basic discoveries to useful products by drawing upon the skills and local knowledge promoted within Dr. Yanti Aesthetic Clinics.

GSCG group managing director, Benito Novas, provided a clear description of the new strategic direction and objectives. "Our goal is to make regenerative medicine benefits a reality for both doctors and patients all around the world. We recently launched a very similar effort in Pakistan. Additional announcements are planned in the near future as we attempt to expand our presence." Meso Numismatics and Global Stem Cells Group Expand its Global Footprint

The current market outlook.

Stem cell therapy is striving to become an increasingly effective clinical solution to treat conditions that traditional or mainstream medicine offers only within palliative care and pain management. Patients all over the world are searching for a natural regenerative alternative without the potential risks and side effects sometimes associated with mainstream pharmaceuticals. With the opening of each new treatment center in populous regions such as Indonesia, GSCG is working to help stem cell therapy and regenerative medicine to eventually move from alternative and elective procedures to mainstream protocols.

This new clinic effort will play a significant role in the development of regenerative medicine in Indonesia and indeed the rest of the world by adding yet another opportunity for continuous improvement through research and development, Christensen continued. By adding busy clinics in population centers, we plan to consistently generate high volumes of reliable clinical data to assist us with the development and refinement of even more medicines and treatments.

About Dr. Yanti Aesthetic Clinics

Dr. Yanti Aesthetic Clinics is a premier cosmetic and aesthetics clinic based in Kelapa Gading, Jakarta Utara. Since its inception in 2004 in Surabaya by Dr. Khoe Yanti Khusmiran, the clinic has expanded to over 6 branches throughout Indonesia. Dr. Yanti clinics provide a range of skin and body enhancement treatments through minimally invasive and non-invasive procedures the expertise of which are a natural fit for the addition of a variety of stem cell therapies.

"Indonesians have a growing need for the latest medical technology that is reliable, potent, has reduced side effects, and leverages the bodys own healing biochemistry to resolve injury and aging, said Dr. Yanti. We are honored to be a part of GSCG, which has a proven 10-year track record in the market with a strong and growing international reputation. This new partnership is expected to create a wide variety of custom treatment options we can offer our patients and treat injury and illness in ways we could not before.

The newly formed partnership will deliver revolutionary medicines through Dr. Yanti clinics to assist patients in avoiding permanent harm and live a healthier life, while changing the paradigm from asymptomatic treatments to cures that may improve and restore quality of life.

More about Global Stem Cells Group

GSCG delivers leadership in regenerative medicine research, patient applications, and training through our strategic global networks. We endeavor to enable physicians to treat otherwise incurable diseases using stem cell therapy and to improve the quality of life and care across the world.

For this reason, GSCG works with innovative, next-generation therapy providers like Dr. Yanti Aesthetic Clinics to give access to one-of-a-kind holistic and safe treatment options.

More information regarding this transaction and the Global Stem Cells Group may be found at GSCG.

This press release should be read in conjunction with all other filings on http://www.sec.gov

For more information on Global Stem Cells Group please visit: http://www.stemcellsgroup.com

About Meso Numismatics: Meso Numismatics, Corp is an emerging Biotechnology and numismatic technology company. The Company has quickly become the central hub for rare, exquisite, and valuable inventory for not only the Meso region, but for exceptional items from around the world.

Meso has now added Biotechnology to its portfolio and will continue to grow the company in this new direction. With the Company's breadth of business experience and technology team, the Company will continue to help companies grow.

Forward-Looking Statements

Some information in this document constitutes forward-looking statements or statements which may be deemed or construed to be forward-looking statements, such as the closing of the share exchange agreement. The words plan, "forecast", "anticipates", "estimate", "project", "intend", "expect", "should", "believe", and similar expressions are intended to identify forward-looking statements. These forward-looking statements involve, and are subject to known and unknown risks, uncertainties and other factors which could cause the Company's actual results, performance (financial or operating) or achievements to differ from the future results, performance (financial or operating) or achievements expressed or implied by such forward-looking statements. The risks, uncertainties and other factors are more fully discussed in the Company's filings with the U.S. Securities and Exchange Commission. All forward-looking statements attributable to Meso Numismatics, Inc., herein are expressly qualified in their entirety by the above-mentioned cautionary statement. Meso Numismatics, Inc. disclaims any obligation to update forward-looking statements contained in this estimate, except as may be required by law.

For further information, please contact:investor.relations@mssvinc.com Telephone: (800) 956-3935

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Global Stem Cells Group Expands Its Stem Cell Therapy and Regenerative Medicine Centers to Indonesia - GlobeNewswire

Skin Stem Cells Comments Off on Global Stem Cells Group Expands Its Stem Cell Therapy and Regenerative Medicine Centers to Indonesia – GlobeNewswire | August 2nd, 2022

Cell Culture Media, Sera, and Reagents Market: Introduction

According to the report, the globalcell culture media, sera, and reagents marketwas valued at US$6.1 Bnin 2020 and is projected to expand at a CAGR of10.3%from 2021 to 2031. Cell culture media, also known as growth media, is an umbrella term that encompasses any gel or liquid created to support cellular growth in an artificial environment. It is a combination of compounds and nutrients designed to support cellular growth.

Cell culture reagents include cell culture media, media supplements, and sterile reagents. Common cell culture reagents are antibiotics and amino acid supplements. Serum is a key component for growing and maintaining cells in culture. It contains a mixture of proteins, hormones, minerals, and other growth factors. It is added to media as a growth supplement, and specialized forms can be used for different experimental conditions.

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Increase in Demand for Cost-effective and Highly Efficient Cell Culture Products to Drive Global Market

Cell culture technology is applied in various domains such as research, academics, bioprocessing & manufacturing, cell therapy, and regenerative medicines. Leading pharmaceutical companies are expanding their capabilities into biopharmaceutical manufacturing in order to leverage high market potential and due to increase in demand for these products.

Rise in demand for cost-effective and highly efficient cell culture products such as bioreactors, media, reagents, and sera for the production of high-yield cell lines has led to an increase in the number of new product launches. This factor is anticipated to provide lucrative opportunities in the global cell culture market during the forecast period.

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Contract Research & Manufacturing and Focus on Stem Cell Research to Propel Market

The cell culture media, sera, and reagents market is witnessing a shift toward contract manufacturing & research, primarily due to significant capital investment and specificity of each biomanufacturing process. For instance, cell cultures could be 2D, 3D, rotating, continuously stirred, batch-fed, and several other types. The expanding scope of cell culture into areas such as stem cell research is boosts the growth of the global market. Rise in importance of stem cell therapy is underlined by the fact that these therapies help treat the cause of the disease, while conventional treatment methods help in managing only the symptoms. This requires advanced capabilities in terms of capital, equipment, and resources; hence, contract manufacturing presents an economically beneficial solution.

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Major Players in Global Cell Culture Media, Sera, and Reagents Market

Key players operating in the global cell culture media, sera, and reagents market include Thermo Fisher Scientific, Inc., Merck KGaA, Cytiva (Danaher Corporation), Becton, Dickinson and Company, Corning Incorporated, HiMedia Laboratories, FUJIFILM Irvine Scientific, Inc., InvivoGen, SeraCare (LGC Clinical Diagnostics, Inc.), and Lonza. Each of these players has been profiled in the cell culture media, sera, and reagents market report based on parameters such as company overview, financial overview, business strategies, application portfolio, business segments, and recent developments.

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The Role of Cell Culture Media, Sera, and Reagents Market Industry Growth, Competitors Analysis, New Technology, Trends and Forecast 2021 2031 -...

Skin Stem Cells Comments Off on The Role of Cell Culture Media, Sera, and Reagents Market Industry Growth, Competitors Analysis, New Technology, Trends and Forecast 2021 2031 -… | August 2nd, 2022

WORCESTER, Mass., July 27, 2022 (GLOBE NEWSWIRE) -- Mustang Bio, Inc.(Mustang) (NASDAQ: MBIO), a clinical-stage biopharmaceutical company focused on translating todays medical breakthroughs in cell and gene therapies into potential cures for hematologic cancers, solid tumors and rare genetic diseases, today announced that the first patient successfully received LV-RAG1 ex vivo lentiviral gene therapy to treat recombinase-activating gene-1 (RAG1) severe combined immunodeficiency (RAG1-SCID), in an ongoing Phase 1/2 multicenter clinical trial taking place in Europe. LV-RAG1 is exclusively licensed by Mustang for the development of MB-110, a first-in-class ex vivo lentiviral gene therapy for the treatment of RAG1-SCID.

Patients with SCID have mutations in blood stem cell genes that are responsible for the development and function of infection-fighting immune cells. As a result, they are unable to mount a normal defense response against infections. The administration of LV-RAG1 includes reduced intensity conditioning prior to reinfusion of the patients own gene-modified blood stem cells.

The patient was administered LV-RAG1 without any complications. LV-RAG1 allowed the patients body to create a functioning immune system, which is responding well to the standard vaccinations for newborns, said Arjan Lankester, Principal Investigator and Professor of Pediatrics and Stem Cell Transplantation at Leiden University Medical Centre (LUMC).

Manuel Litchman, M.D., President and Chief Executive Officer of Mustang said, This first successful administration to a RAG1-SCID patient of a stem-cell based gene therapy represents a significant positive step forward for our MB-110 development program. This treatment, along with our X-linked severe combined immunodeficiency (XSCID) programs, which includes MB-107 and MB-207, has established Mustang as a leader in developing treatments for SCID patients, who are in great need of these life-saving therapies. XSCID and RAG1-SCID make up almost 60% of all SCID cases combined.1 We look forward to continuing to advance these clinical candidates, including plans to initiate a multicenter pivotal Phase 2 trial for MB-107 under Mustangs IND in the second half of this year.

LV-RAG1 has been granted Orphan Drug Designation by the European Medicines Agency. Additional clinical trial sites are expected to be added in the near future.

Signed in 2021, Mustangs exclusive, worldwide license agreement for LV-RAG1 established an ongoing partnership with LUMC and LUMCs Frank J. Staal, Ph.D., molecular immunologist and professor of Molecular Stem Cell Biology. The license agreement grants Mustang rights to certain additional lentiviral gene therapies being developed in Dr. Staals lab.

About RAG1-SCIDSevere combined immunodeficiency (SCID) due to complete RAG1 deficiency is a rare, genetic severe combined immunodeficiency disorder caused by null mutations in the RAG1 gene resulting in less than 1% of wild type V(D)J recombination activity. Patients present with neonatal onset of life-threatening, severe, recurrent infections by opportunistic fungal, viral and bacterial micro-organisms, as well as skin rashes, chronic diarrhea, failure to thrive and fever. Immunologic observations include profound T- and B-cell lymphopenia, low or absent serum immunoglobulins, and normal natural killer cell counts. As is the case with other types of SCID, RAG1-SCID is fatal in infancy unless immune reconstitution is achieved with allogeneic hematopoietic stem cell transplantation (HSCT), or autologous stem cells corrected by gene therapy.

About MB-110 (Ex Vivo Lentiviral Gene Therapy)MB-110 is a first-in-class ex vivo lentiviral gene therapy under development to treat RAG1-SCID, utilizing the LV-RAG1 vector developed in the laboratory of Frank J. Staal, Ph.D., molecular immunologist and professor of Molecular Stem Cell Biology at LUMC. Exclusively licensed to Mustang in 2021, LV-RAG1 is currently being evaluated in a Phase 1/2 multicenter, academic clinical trial (RECOMB) in Europe. Additional information on the trial can be found at http://www.clinicaltrials.gov using the identifier NCT04797260.

The same lentiviral vector drug substance produced by LUMC will be used to transduce patients cells to create the MB-110 drug product produced at Mustang Bios Worcester, MA, cell processing facility for further clinical development and to facilitate eventual commercial launch of the product.

About Mustang BioMustang Bio, Inc. is a clinical-stage biopharmaceutical company focused on translating todays medical breakthroughs in cell and gene therapies into potential cures for hematologic cancers, solid tumors and rare genetic diseases. Mustang aims to acquire rights to these technologies by licensing or otherwise acquiring an ownership interest, to fund research and development, and to outlicense or bring the technologies to market. Mustang has partnered with top medical institutions to advance the development of CAR T therapies across multiple cancers, as well as lentiviral gene therapies for severe combined immunodeficiency. Mustang is registered under the Securities Exchange Act of 1934, as amended, and files periodic reports with the U.S. Securities and Exchange Commission (SEC). Mustang was founded by Fortress Biotech, Inc. (NASDAQ: FBIO). For more information, visit http://www.mustangbio.com.

ForwardLooking Statements This press release contains forward-looking statements within the meaning of Section 27A of the Securities Act of 1933 and Section 21E of the Securities Exchange Act of 1934, each as amended. Such statements, which are often indicated by terms such as anticipate, believe, could, estimate, expect, goal, intend, look forward to, may, plan, potential, predict, project, should, will, would and similar expressions, include, but are not limited to, any statements relating to our growth strategy and product development programs, including the timing of and our ability to make regulatory filings such as INDs and other applications and to obtain regulatory approvals for our product candidates, statements concerning the potential of therapies and product candidates, and any other statements that are not historical facts. Forward-looking statements are based on managements current expectations and are subject to risks and uncertainties that could negatively affect our business, operating results, financial condition and stock value. Factors that could cause actual results to differ materially from those currently anticipated include: risks relating to our growth strategy; our ability to obtain, perform under, and maintain financing and strategic agreements and relationships; risks relating to the results of research and development activities; risks relating to the timing of starting and completing clinical trials; uncertainties relating to preclinical and clinical testing; our dependence on third-party suppliers; our ability to attract, integrate and retain key personnel; the early stage of products under development; our need for substantial additional funds; government regulation; patent and intellectual property matters; competition; as well as other risks described in Part I, Item 1A, Risk Factors, in our Annual Report on Form 10-K filed on March 23, 2022, subsequent Reports on Form 10-Q, and our other filings we make with the SEC. We expressly disclaim any obligation or undertaking to release publicly any updates or revisions to any forward-looking statements contained herein to reflect any change in our expectations or any changes in events, conditions or circumstances on which any such statement is based, except as required by law, and we claim the protection of the safe harbor for forward-looking statements contained in the Private Securities Litigation Reform Act of 1995.

Company Contacts:Jaclyn Jaffe and Bill BegienMustang Bio, Inc.(781) 652-4500ir@mustangbio.com

Investor Relations Contact:Daniel FerryLifeSci Advisors, LLC(617) 430-7576daniel@lifesciadvisors.com

Media Relations Contact:Tony Plohoros6 Degrees(908) 591-2839tplohoros@6degreespr.com

1 Fischer A, et al. Nat Rev Dis Primers. 2015; article number 15061; doi: 10.1038/nrdp.2015.61

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Mustang Bio Announces First Patient Successfully Treated by Ex Vivo Lentiviral Gene Therapy to Treat RAG1 Severe Combined Immunodeficiency - BioSpace

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Autologous Cell Therapy Market 2021-2025: Scope

The autologous cell therapy market report covers the following areas:

Autologous Cell Therapy Market 2021-2025: Segmentation

Learn about the contribution of each segment summarized in concise infographics and thorough descriptions. View a PDF Sample Report

Autologous Cell Therapy Market 2021-2025: Vendor Analysis

We provide a detailed analysis of around 25 vendors operating in the autologous cell therapy market, including Bayer AG, Brainstorm Cell Therapeutics Inc., Daiichi Sankyo Co. Ltd., FUJIFILM Holdings Corp., Holostem Terapie Avanzate Srl, Osiris Therapeutics Inc., Takeda Pharmaceutical Co. Ltd., Teva Pharmaceutical Industries Ltd., Sumitomo Chemical Co. Ltd., and Vericel Corp. among others. The key offerings of some of these vendors are listed below:

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Autologous Cell Therapy Market 2021-2025: Key Highlights

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Autologous Cell Therapy Market Scope

Report Coverage

Details

Page number

120

Base year

2020

Forecast period

2021-2025

Growth momentum & CAGR

Accelerate at a CAGR of 14.16%

Market growth 2021-2025

USD 4.11 billion

Market structure

Fragmented

YoY growth (%)

13.5

Regional analysis

North America, Europe, APAC, and South America

Performing market contribution

North America at 43%

Key consumer countries

US, UK, Germany, Canada, and Japan

Competitive landscape

Leading companies, competitive strategies, consumer engagement scope

Companies profiled

Bayer AG, Brainstorm Cell Therapeutics Inc., Daiichi Sankyo Co. Ltd., FUJIFILM Holdings Corp., Holostem Terapie Avanzate Srl, Osiris Therapeutics Inc., Takeda Pharmaceutical Co. Ltd., Teva Pharmaceutical Industries Ltd., Sumitomo Chemical Co. Ltd., and Vericel Corp.

Market Dynamics

Parent market analysis, market growth inducers and obstacles, fast-growing and slow-growing segment analysis, COVID-19 impact and future consumer dynamics, market condition analysis for the forecast period

Customization purview

If our report has not included the data that you are looking for, you can reach out to our analysts and get segments customized.

Table Of Contents :

Executive Summary

Market Landscape

Market Sizing

Five Forces Analysis

Market Segmentation by Product

Customer landscape

Geographic Landscape

Vendor Landscape

Vendor Analysis

Appendix

About Us

Technavio is a leading global technology research and advisory company. Their research and analysis focus on emerging market trends and provide actionable insights to help businesses identify market opportunities and develop effective strategies to optimize their market positions. With over 500 specialized analysts, Technavio's report library consists of more than 17,000 reports and counting, covering 800 technologies, spanning across 50 countries. Their client base consists of enterprises of all sizes, including more than 100 Fortune 500 companies. This growing client base relies on Technavio's comprehensive coverage, extensive research, and actionable market insights to identify opportunities in existing and potential markets and assess their competitive positions within changing market scenarios.

Contact

Technavio ResearchJesse MaidaMedia & Marketing ExecutiveUS: +1 844 364 1100UK: +44 203 893 3200Email: [emailprotected]Website: http://www.technavio.com/

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Autologous Cell Therapy Market Size to Grow by USD 4.11 billion, Bayer AG and Brainstorm Cell Therapeutics Inc. Among Key Vendors - Technavio - PR...

Cardiac Stem Cells Comments Off on Autologous Cell Therapy Market Size to Grow by USD 4.11 billion, Bayer AG and Brainstorm Cell Therapeutics Inc. Among Key Vendors – Technavio – PR… | August 2nd, 2022

Dr. Jennifer Lang splits most of her work life treating patients at Gates Vascular Institute and conducting research in her lab several floors up in the same building.

UB medical physics students Simon Wu and Emily Vanderbelt work with flow-through 3D-printed aneurysm models using X-rays in the Canon Stroke & Vascular Research Center, part of the University at BuffaloClinical and Translational Research Center on the Buffalo Niagara Medical Campus.

The arrangement suits her well as she continues promising research to learn if a stem cell-derived treatment can repair damaged heart tissue.

Lang, a cardiologist, and her University at Buffalo team, face a dilemma: The immune system revs into high gear when the heart suffers a serious setback, limiting the power of stem cells to heal.

The daunting task seems more surmountable these days because she works in a building filled with researchers of all stripes.

I do collaborations with groups that I otherwise wouldn't have. Its led to some really new, interesting results, said Lang, assistant professor in the UB Jacobs School of Medicine and Biomedical Sciences who practices with UBMD Internal Medicine and at the Buffalo VA Medical Center.

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This day, a surgical team worked seamlessly to monitor her vital signs and feather a medical device through a catheter into the left side of her damaged heart. The procedure slowed her heartrate so her organs could take a couple of days to re-collect themselves and give her a fighting chance to recover.

UB-fueled research unfolds on floors five through eight of the building at 875 Ellicott St., alongside Buffalo General Medical Center.

Ten years ago, the university invested $118 million into its Clinical and Translational Research Center, and about $25 million for equipment came from industry partners who wanted to join forces with physicians, engineers and others in the science fields.

The center became the first major pieceof the UB medical school to move onto the downtown Buffalo Niagara Medical Campus, followed in late 2017 by the $375 million Jacobs School teaching and research complex, around the corner at Main and High streets.

Both foster translational medicine, which combines disciplines, resources and techniques to move benchtop research to the patient bedside, eventually strengthening community health.

Langs work symbolizes the approach.

The Buffalo native can see her high school alma mater, City Honors, from her workplace. She went to Cornell University as an undergraduate and returned to Buffalo to go to medical school. Buoyed by fellow UB students, faculty and mentors, she chose to stay in the city for her internal medicine residency and cardiology fellowship.

Lang did her classroom work and research on the UB South Campus and most of her clinical work 8 miles away, on the downtown Medical Campus.

Stairs and elevators are the only things that separate her from most of her collaborators and patients today.

I moved into this building when it opened 10 years ago, she said. At the time, I was completing my cardiology fellowship. There was a physical divide, so I was thrilled with the new arrangement. Things can happen in parallel now.

Dr. Timothy Murphy, left, director of theUB Clinical and Translational Research Center in Buffalo, works with research technician Charmaine Kirkham in their lab, which focuses on potential treatments forchronic obstructive pulmonary disease (COPD).

That was the plan, said Dr. Timothy Murphy, director of the UB Clinical and Translational Research Center.

Clinical research and health care have become more and more seamlessly integrated, he said. The building contributed to that.

Murphy, another regional native, was among those who shared and helped carry out the vision of Gates Vascular Institute founder Dr. L. Nelson Nick Hopkins III, who chaired the UB Department of Neurosurgery from 1989 to 2013 and wanted to create a more innovative vascular center.

Murphy moved his lab in 2006 from the VA Medical Center near South Campus to the UB Center for Bioinformatics and Life Sciences on the Medical Campus, so he could be involved in the design of the UB research center, on floors above Gates Vascular, as well as at the Jacobs School particularly its labs.

They always talked about physicians and researchers bumping into each other, talking to each other, and having graduate students and postdocs and technicians talk to each other, Murphy said. Having done it now for all these years, I see it really does work.

He and his research team continue a 20-year study on the bacterial infection that causes COPD in hopes it will help lead to vaccines that prevent the infection and new treatments to clear the bacteria from the lower airway.

As senior associate dean forclinical and translational researchat the Jacobs School, he is also the point person for coordinating UB-related clinical trials and encouraging collisions between health care researchers on the Medical Campus and around the world.

There were 70 such trials on the Medical Campus in 2015, when the building where he works was in its infancy. Today, there are more than 200.

"Things can happen in parallel now," says Dr. Jennifer Lang, a cardiologist, researcher and University at Buffalo assistant professor who splits her research and clinical time in the same building on the Buffalo Niagara Medical Campus.

Labs focused on obstetric and gynecological advances and keys to healthy aging occupy space near his seventh-floor lab.

The Clinical and Translational Research Center was established in 2012. UB added a biobank in 2019 to store medical specimens for ongoing clinical studies.

Its collaborative framework helped UB land a $15 million Clinical and Translational Science Awardin 2015 from the National Institutes of Health (NIH) to encourage research efforts across university departments and specialties to boost innovation, speed development of medical treatments, and reduce health disparities in poor, rural and minority communities.

The five-year grant was renewed in 2020 with nearly $22 million more, encouraging Buffalo-based researchers to work with others who got awards, including researchers with Harvard, Johns Hopkins, Stanford and Yale universities.

A printer creates a 3D model, slice by slice, at the Canon Stroke & Vascular Research Center in the University at Buffalo Clinical and Translational Research Center. Lab researchers experiment with different mixtures of six polymers to make the most malleable and useful models for medical research.

Throughout the building, the goal is to improve medical devices and treatments that make an impact in the clinics and catheter suites in the Gates Vascular Institute on the floors below the research center and provide data and education that informs others, including patients.

The eighth-floor Canon Stroke & Vascular Research Center, which tops the UB research center, is a case in point.

Ciprian Chip Ionita, its director, came to UB from Romania in 1999 and worked his first dozen years on the South Campus.

We were the first ones to move in, said Ionita, assistant professor of biomedical engineering and member of the medical school's Department of Neurosurgery.

The lab was designed to help innovate and improve medical devices and neurovascular procedures.

Part of its work involves using MRIs, CT scans and other radiological images of Gates Vascular patients to create 3D-printed models of the circulatory system and heart.

3D printing created this replica of part of a patient's spinal column at the Canon Stroke & Vascular Research Center. Researchers there push the boundaries until their findings are refined to the point where they can be applied to model-making on two highly calibrated 3D printers in the Jacobs Institute downstairs from the lab that meet FDA standards. We fail up here about 90% of the time, says Ciprian Chip Ionita, lab director. They fail maybe 1%, so were testing everything that's possible.

Medical school and other lab researchers use the models produced here to better understand how anatomy and disease of former and current patients led to poor health and, in some cases, poor surgical outcomes.

Gates Vascular surgeons also can use 3D models that replicate the anatomy of patients awaiting surgery to practice feathering catheters and medical devices through bends, nooks and crannies of the blood vessels, and deploy medical devices in spines and the circulatory system as they maneuver past muscles, bones, blockages and other obstructions that might come into play.

During practice interventions, we analyze everything, Ionita said, because we can go into these models with sensors to measure blood flow, blood pressure and more.

You can create a model that says, Here's somebody who has a carotid artery that's 50% (blocked) and he's 50 years old, Ionita said. Or we can say, 'Here is a young person in their 20s, and is fully compliant, no stenosis or whatever.' And those mechanical properties are translated by the printer.

Even cadaver donors cant do that.

The goal is to lower the rate of complications and be successful in one shot during a procedure, said Ionita, who supervises up to 10 graduate biomedical engineering students, and roughly 20 undergraduate, graduate and medical school students.

Those who pay close attention to 3D models and other medical research based on data from patients treated in the building include Dr. Elad Levy, co-director of the Gates Vascular Stroke Center; Dr. Adnan Siddiqui, director of neurological and stroke services at Kaleida Health; and Dr. Vijay Iyer, medical director of cardiology and the Structural Heart Program at Kaleida. All three have ties to UB.

Even here, Ionita said, physician-scientists and other researchers see the damage that smoking, high blood pressure and living in ZIP codes where poverty is rampant can create complications that lead to worse health and surgical outcomes.

Eric Wozniak, a senior research and development technician in the Idea to Reality lab at the Jacobs Institute, uses a microscope as he works to improve catheter technology.

Doctors and staff improve treatment protocols and surgical prowess with help from those who work on the top half of the building for UB and the Jacobs Institute. The latter is named for Dr. Lawrence D. Jacobs, a Buffalo neurosurgeon whose research led to the first treatments for multiple sclerosis.

Four years after Jacobs died in 2001, his brother Jeremy, chair of the Delaware North Cos. and the UB Council, approached the university about creating a lasting memorial for the respected physician. He later signed on to the concept of creating a multidisciplinary vascular center, starting with a $10 million donation for the institute that bears the family name.

The institute includes an atrium, caf and glass-walled spaces that overlook procedure rooms on the floor below. It has 50 employees, including more than 30 biomedical and electrical engineers, who seek company-sponsored research funding, help collect data and make prototypes for clinical trials, and work with researchers to publish their work in medical journals.

In 2016, the institute was designated a 3D Printing Center of Excellence in Health Care by Israeli-based Stratasys Ltd., a leading 3D printing-maker. In early 2018, it created a proof-of-concept Idea to Reality Center, known as i2R, to further improve medical devices and surgical techniques in the vascular space.

This is our secret sauce lab, said Siddiqui, Jacobs Institute CEO. There's nothing we do downstairs that we could not do better.

This is a device designed and built in the Idea 2 Reality lab at the Jacobs Institute in Buffalo. The lab improves medical devices and technology used in vascular procedures and treatments.

Dr. Carlos Pena, who ran the FDA Neurologic Devices Division for 15 years, joined the institute staff last year to improve the chances technology conceived and designed with help from the institute gets to market.

Every company wants to talk to him, Siddiqui said. He tells them what testing needs to be done. Some of that gets done in-house. A lot of it goes to the university or, when they have a clinical trial, that gets done downstairs so the entire ecosystem is functioning, I think better than Nick Hopkins ever imagined.

Lang, the cardiologist, doesnt miss her former workday commutes. She loves the design and location of the building that sets the standard for vascular care.

Most of her days mix benchtop research in her lab and patient visits and procedures on the floors below. When there is time, she can visit her husband, Fraser Sim, neuroscience director and associate professor at the medical school.

Because we're in such close proximity to the Jacobs School now, we're also really able to engage the medical students earlier in their careers and encourage more research, Lang said. And because we're so close to the hospital, we're able to involve medical residents and fellows in our research projects much more than ever before.

University at Buffalo medical school postdoctoral research associateToubaTarvirdizadeh focuses on cardiac research in the lab of Dr. Jennifer Lang at the UB Clinical and Translational Research Center in Buffalo.

She has spent a decade trying to find better ways for a stem cell derivative that can withstand an immune response and rejuvenate heart tissue without major complications, a result that could help patients recover from a heart attack and lessen the strain of heart failure.

Four years ago, Lang and her doctoral student researcher, Kyle Mentkowski, discovered a way that lowered the immune response in mice that received the derivative.

Mentkowski, now a post-doctorate researcher at Harvard-affiliated Massachusetts General Hospital, was talking with another group of student researchers in the building when they thought it might be a good idea to bring Dr. Jessica Reynolds, an immunologist and UB medical school associate professor, into the research.

The collaboration created robust, reproducible results in mice models, Lang said, and the start of testing in human immune cells she and her colleagues hope can benefit patients within the next decade.

Collaborators now regularly get together to chat at the Jacobs Institute.

The NIH seems very interested in this as a potential clinical therapy, Lang said, but the field as a whole is still in the beginning stages of understanding where we need to go next.

Dr. Aaron Hoffman, left, University at Buffalo medical school associate professor of surgery, and Dr. Kenneth Snyder, UB associate professor of neurosurgery, chat during a break in the Jacobs Institute atrium.

UB researchers have shared some of their findings with researchers making similar inroads elsewhere, she said, and the work spawned other collaborations with Reynolds, her research team and scientists in the UB Department of Biomedical Engineering.

This type of unplanned interaction is not a unique occurrence in this building, Lang said. Our story is just one of many.

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Buffalo center fuels research that can save your life from heart disease and stroke - Buffalo News

Cardiac Stem Cells Comments Off on Buffalo center fuels research that can save your life from heart disease and stroke – Buffalo News | August 2nd, 2022

The British Heart Foundation announced the winner of its $36 million Big Beat Challenge, one of the largest non-commercial awards ever given for heart research.

The winning team, CureHeart, brings together researchers from the U.K., U.S. and Asia, including Eric Olson, professor and chair of the Department of Molecular Biology at UT Southwestern Medical Center.

Olson is the founding chair of the department and directs the Hamon Center for Regenerative Science and Medicine and the Wellstone Center for Muscular Dystrophy Research. He holds the Robert A. Welch Distinguished Chair in Science and the Annie and Willie Nelson Professorship in Stem Cell Research.

He has spent his career investigating heart and muscle development and disease, leading to his participation on the CureHeart team. The Olson Lab at UTSW has been incredibly successful in muscular research, most recently providing a new way to correct the mutation that causes Duchenne muscular dystrophy through gene editing.

CureHeart made the top of the list with its gene editing therapy aimed at curing inherited heart muscle diseases, known as cardiomyopathies.

A BHF release said the technology will seek to develop the first cures for inherited heart muscle diseases by pioneering revolutionary and ultra-precise gene therapy technologies that could edit or silence the faulty genes that cause these deadly conditions.

The project will use gene-editing technology CRISPR to complete base and prime editing in the heart for the first time.

It works by correcting or silencing a faulty gene in the pumping machinery of the heart, either by re-writing the DNA at a single location or by switching off the entire copy of the faulty gene.

The technique was described as molecules that act like tiny pencils to rewrite the single mutations that are buried within the DNA of heart cells in people with heart conditions.

It can also help the heart produce enough proteins to function normally, again by fixing or stimulating the faulty gene.

With ultra-precise base editing technology, we hope to be able to correct a single letter and larger errors in the genetic code. This would mark a breakthrough for not only genetic cardiomyopathies, but for many heart conditions, said Olson in the release.

The project is the next step toward a real-world application, having already proved successful in animals with cardiomyopathies and in human cells. Members of the team believe therapies could be delivered through an arm injection, slowing or stopping the progression of cardiomyopathies, or even curing the disease entirely.

If successful, the research could have enormous impacts.

Every year in the US, around 2,000 people under the age of 25 die of a sudden cardiac arrest, often caused by one of these inherited muscle diseases, said the release. Current treatments do not prevent the condition from progressing, and around half of all heart transplants are needed because of cardiomyopathy.

The researchers believe it could also be successful in preventing the disease from being expressed if inherited. Children who receive the faulty gene from their parents could receive the injection and never develop cardiomyopathy in the first place.

Over the last 30 years, we have made extraordinary advancements in our understanding of the genetic mistakes that cause cardiomyopathy. CureHeart is a once-in-a-generation opportunity to transform this knowledge into a cure, said Olson in the release.

The technology is still in the research and development phase, but Olson said the funds will be used to optimize the method and expand it to a larger number of genetic diseases of the heart, and could move to clinical trials in the next few years.

See the article here:
UTSW researcher part of team awarded $36 million heart research grant - The Dallas Morning News

Cardiac Stem Cells Comments Off on UTSW researcher part of team awarded $36 million heart research grant – The Dallas Morning News | August 2nd, 2022

image:Differentiation of pluripotent stem cells (PSCs) is regulated through a methionine-mediated mechanism, which has now been pinpointed by Tokyo Tech researchers. They have revealed that zinc (Zn) plays a crucial role in PSC potentiation. They used these insights to design a protocol to convert PSCs into insulin-producing pancreatic cellsa high-potential diabetes therapy. view more

Credit: Prof. Shoen Kume from Tokyo Institute of Technology

Differentiation of pluripotent stem cells (PSCs) is regulated through a methionine-mediated mechanism, which has now been pinpointed by Tokyo Tech researchers. They have revealed that zinc (Zn) plays a crucial role in PSC potentiation. They used these insights to design a protocol to convert PSCs into insulin-producing pancreatic cellsa high-potential diabetes therapy.

Stem cell research has gained a lot of attention in the world of medical therapeutics. Pluripotent stem cells (PSCs) can self-renew and transform into different types of cells in the body via a process called differentiation. These cells have manifold applications, such as disease modeling, drug discovery, and cell replacement therapy.

One area of focus in PSC research is diabetes treatments. A common characteristic of diabetes is having ineffective or overworked pancreatic cellscells that produce insulin. Controlling the differentiation of PSCs to produce cells is one of the major goals of research in the field. Previous studies have shown that methionine, an amino acid, plays a major role in the differentiation of PSCs. But the precise mechanism behind this has been, thus far, unknown.

To find the missing piece of this puzzle, a team of researchers from Japan, led by Prof. Shoen Kume from Tokyo Institute of Technology (Tokyo Tech), delved deeper into the methionine-mediated regulation of PSC pluripotency. In a recent study published in Cell Reports, the researchers revealed that cellular zinc (Zn) content played a crucial role in stem cell differentiation. Prof. Kume explains, Earlier research in the area has shown that if we culture PSCs in a medium which is deficient in methionine, it leads to a reduction in intracellular S-adenosyl methionine or SAM, which renders PSCs in a state of potentiated differentiation. But our study further identified that zinc (Zn) is a downstream target of methionine metabolism and it can potentiate pluripotency in undifferentiated PSCs.

In this study, the research team first cultured PSCs in a methionine-deprived environment. They found that methionine-deprivation not only reduced the intracellular protein-bound Zn levels in cells, but that it also upregulated SLC30A1, a gene that produces an important Zn transport protein.

The team then cultured hiPSCs under low Zn concentrations. They discovered that a Zn-deprived medium partially mimicked methionine deprivation and led to a decrease in cell growth and an increase in potentiated differentiation. They also found that the Zn deprived state also altered the methionine metabolism profile and eliminated undifferentiated hiPSCs. These results indicated that methionine deprivation-induced differentiation takes place by lowering the Zn content in cells.

Using the insights, the team then developed a methodology for generating insulin-producing pancreatic cells. cell transplantation is a promising treatment for diabetes, but there is a paucity of donor cells for the treatment, as well as immune-related complications that can arise from this treatment. Using PSCs to produce genetically-matching cells is a way to overcome this, explains Prof. Kume.

These findings indicate a link between Zn mobilization and methionine-induced potentiation of PSCs and provide clear a direction for future research in the field of stem cell therapies.

Related Information

Today's Stem Cell Special: Small Intestine on a Plate! https://www.titech.ac.jp/english/news/2021/048927

A Ferry Protein in the Pancreas Protects It from the Stress Induced by a High-Fat Diet | Tokyo Tech Newshttps://www.titech.ac.jp/english/news/2020/047867.html

Move over Akita: Introducing 'Kuma Mutant' Mice for Islet Transplantation Researchhttps://www.titech.ac.jp/english/news/2020/047462

Shoen Kume - Towards a new therapy for diabetes - Regenerating pancreas from ES and iPS cellshttps://www.titech.ac.jp/english/public-relations/research/stories/faces37-kume

Kume &Shiraki Lab.http://www.stem.bio.titech.ac.jp/index.html

About Tokyo Institute of Technology

Tokyo Tech stands at the forefront of research and higher education as the leading university for science and technology in Japan. Tokyo Tech researchers excel in fields ranging from materials science to biology, computer science, and physics. Founded in 1881, Tokyo Tech hosts over 10,000 undergraduate and graduate students per year, who develop into scientific leaders and some of the most sought-after engineers in the industry. Embodying the Japanese philosophy of monotsukuri, meaning technical ingenuity and innovation, the Tokyo Tech community strives to contribute to society through high-impact research.

https://www.titech.ac.jp/english/

Experimental study

Cells

Methionine metabolism regulates pluripotent stem cell pluripotency and differentiation through zinc mobilization

19-Jul-2022

Disclaimer: AAAS and EurekAlert! are not responsible for the accuracy of news releases posted to EurekAlert! by contributing institutions or for the use of any information through the EurekAlert system.

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The zinc link: Unraveling the mechanism of methionine-mediated pluripotency regulation - EurekAlert

IPS Cell Therapy Comments Off on The zinc link: Unraveling the mechanism of methionine-mediated pluripotency regulation – EurekAlert | July 25th, 2022

The company is developing a novel, non-invasive, bio-guided treatment to restore function of patients with acute spinal cord injuries.

Over two hundred and fifty thousand people suffer from spinal cord injuries in the US every year, with patients typically experiencing major, and mostly irreversible, loss of function that requires millions of dollars in lifetime care per patient.

NurExone is developing a revolutionary bio-guided treatment. The technology is based on exosomes, small particles that are created when stem cells proliferate, to deliver therapeutic agents to a specific location in the body. Nurexones proprietary agents, delivered by the exosomes, create an environment may support Nerves regeneration. For spine injuries, the bio-guided treatment is an agent that inhibits the PTEN protein in nerve cells, allowing nerves regeneration to occur.

The company carried out preclinical, animal studies that demonstrated that bio-guided treatment led to significant improvement, sensory recovery, and faster reflex restoration. The study reveals that Nurexones proprietary technology caused new connections in the spinal cord, repairing the damage from injuries, at least in part.

Studies also suggested that Nurexones technology may be useful for other indications including strokes and traumatic brain injuries (TBI).

The company was founded in 2020, based on research by Professor Shulamit Levenberg, Head of the Biomedical Engineering Department at Technion, and by Professor Daniel Offen, Head of the Lab for Neurosciences at the Felsenstein Medical Research Center in Tel Aviv University.

Spine related injuries are expected to increase in the future owing to motor accidents, workplace injuries, stroke, and cancer related motor disabilities. Currently, between 250,000 and 500,000 people become spinal cord injured every year worldwide, and the lifetime costs of treatments range from $1.6 million to nearly $5 million for 25-year-olds, to $1.1 million to nearly $2.7 million for 50-year-olds. The total addressable market for spinal cord trauma injuries is expected to reach $3.04 billion by 2025, with a CAGR of 3.7%.

Stepping back to look solely at exosome technology (not necessarily related to SCI), since 2018, exosomes are an emerging therapeutic field, with hundreds of millions of US dollars invested in exosome technologies by companies including Eli Lilly, Roche, and Takeda.

NurExone has obtained exclusive rights to an advanced exosome manufacturing process developed at the Technion, Israel Institute of Technology, Haifa. NurExone will be responsible for additional exosome research, management of clinical studies and commercialization of the technology for different indications not limited to Central Nerve System (CNS).

NurExones listing on the TSX.V under the symbol NRX was accomplished through an agreed reverse takeover (RTO) of EnerSpar signed on January 3, 2022. EnerSpar will acquire each ordinary share of NurExone in exchange for 17 post-consolidation EnerSpar shares, resulting in a total of 48,383,963 total shares outstanding following completion of the transaction.

Despite limited financial analyses available on the stock, it seems like a potentially unique opportunity given the fact that the market for spinal-cord treatment continues to grow, thus enabling new players in the field to partake in this ever-growing industry. Moreover, any company that delivers therapy that has the potential to unlock the secret of restoring function to patients who have experienced traumatic spinal injury, seems to be worth considering

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New TSXV listing looks to address the $3B spinal cord injury treatment market (NRX.V) - FXStreet

Spinal Cord Stem Cells Comments Off on New TSXV listing looks to address the $3B spinal cord injury treatment market (NRX.V) – FXStreet | July 25th, 2022

Ethics statement

All human material (blood RNA, primary microglia RNA, iPSCs) used in this study was derived after signed informed consent: for blood, according to University of Oxford OHS policy document 1/03; all procedures related to the use of the primary microglia followed established institutional (McGill University, Montreal, QC, Canada) and Canadian Institutes of Health Research guidelines for the use of human cells; for iPSC, with approval from the South Central Berkshire Research Ethics Committee, U.K. (REC 10/H0505/71). The blood RNA and primary microglia RNA samples have been published previously26, as have the iPSC lines (see below).

Four healthy control iPSC lines, SFC840-03-03 (female, 67years old,35), SFC841-03-01 (male, 36,18), SFC856-03-04 (female, 78,36), OX3-06 (male, 49,37), generated from skin biopsy fibroblasts and characterized as described before, were used in this study. Additionally, the previously reported26 line AH016-3 Lenti_IP_RFP (male, 80years old), which constitutively expresses Red Fluorescent Protein (RFP) under continuous puromycin selection, was used for some live-imaging experiments.

iPSCs were cultured in mTeSR1 (StemCell Technologies) or OXE8 medium38 on Geltrex (Thermo Fisher)-coated tissue culture plates with daily medium changes. Passaging was done as clumps using EDTA in PBS (0.5mM). Cells were initially expanded at low passage to create a master stock, which was used for all experiments to ensure consistency. Cells were regularly tested negative for mycoplasma using MycoAlert Mycoplasma Detection Kit (Lonza).

iPSCs were differentiated to MNs according to our previously published protocol18,19,27. Briefly, neural induction of iPSC monolayers was performed using DMEM-F12/Neurobasal 50:50 medium supplemented with N2 (1X), B27 (1X), 2-Mercaptoethanol (1X), AntibioticAntimycotic (1X, all ThermoFisher), Ascorbic Acid (0.5M), Compound C (1M, both Merck), and Chir99021 (3M, R&D Systems). After two days in culture, Retinoic Acid (RA, 1M, Merck) and Smoothened Agonist (SAG, 500nM, R&D Systems) were additionally added to the medium. Two days later, Compound C and Chir99021 were removed from the medium. After another 5days in culture, neural precursors were dissociated using accutase (ThermoFisher), and split 1:3 onto Geltrex-coated tissue culture plates in medium supplemented with Y-27632 dihydrochloride (10M, R&D Systems). After one day, Y-27632 dihydrochloride was removed from the medium, and then the cells were cultured for another 8days with medium changes every other day. For terminal maturation, the cells were dissociated on day in vitro (DIV) 18 using accutase and plated onto coverslips or tissue culture plates coated with polyethylenimine (PEI, 0.07%, Merck) and Geltrex or tissue culture dishes coated with PDL (Sigma-Aldrich)/ Laminin (R&D Systems)/ Fibronectin (Corning). For this step, the medium was additionally supplemented with BDNF (10ng/mL), GDNF (10ng/mL), Laminin (0.5g/mL, all ThermoFisher), Y-27632 dihydrochloride (10M), and DAPT (10M, R&D Systems). Three days later, Y-27632 dihydrochloride was removed from the medium. After another three days, DAPT was removed from the medium. Full medium changes were then performed every three days.

For MNs differentiated in co-culture medium alone, all steps were performed similarly until three days after the terminal re-plating (D21). MNs were then cultured in co-culture medium as described below.

iPSCs were differentiated to macrophage/microglia precursors as described previously20,21. Briefly, embryoid body (EB) formation was induced by seeding iPSCs into Aggrewell 800 wells (STEMCELL Technologies) in OXE838 or mTeSR1 medium supplemented with Bone Morphogenetic Protein 4 (BMP4, 50ng/mL), Vascular Endothelial Growth Factor (VEGF, 50ng/mL, both Peprotech), and Stem Cell Factor (SCF, 20ng/mL, Miltenyi Biotec). After four days with daily medium changes, EBs were transferred to T175 flasks (~150 EBs each) and differentiated in X-VIVO15 (Lonza), supplemented with Interleukin-3 (IL-3, 25ng/mL, R&D Systems), Macrophage Colony-Stimulating Factor (M-CSF, 100ng/mL), GlutaMAX (1X, both ThermoFisher), and 2-Mercaptoethanol (1X). Fresh medium was added weekly. After approximately one month, precursors emerged into the supernatant and could be harvested weekly. Harvested cells were passed through a cell strainer (40M, Falcon) and either lysed directly for RNA extraction or differentiated to microglia in monoculture or co-culture as described below.

Three days after the final re-plating of differentiating MNs (DIV21), macrophage/microglia precursors were harvested as described above and resuspended in co-culture medium comprised of Advanced DMEM-F12 (ThermoFisher) supplemented with GlutaMAX (1X), N2 (1X), AntibioticAntimycotic (1X), 2-Mercaptoethanol (1X), Interleukin-34 (IL-34, 100ng/mL, Peprotech), BDNF (10ng/mL), GDNF (10ng/mL), and Laminin (0.5g/mL). MNs were quickly rinsed with PBS, and macrophage/microglia precursors re-suspended in co-culture medium were added to each well. Co-cultures were then maintained for at least 14days before assays were conducted as described below. Half medium changes were performed every 23days.

For comparisons between co-cultures and monocultures, MNs and monocultured microglia were also differentiated alone in co-culture medium.

Cells cultured on coverslips were pre-fixed with 2% paraformaldehyde in PBS for 2min and then fixed with 4% paraformaldehyde in PBS for 15min at room temperature (RT). After permeabilization and blocking with 5% donkey/goat serum and 0.2% Triton X-100 in PBS for 1h at RT, the coverslips were incubated with primary antibodies diluted in 1% donkey/goat serum and 0.1% Triton X-100 in PBS at 4C ON. The following primary antibodies were used: rabbit anti-cleaved caspase 3 (1:400, 9661S, Cell Signaling), mouse anti-ISLET1 (1:50, 40.2D6, Developmental Studies Hybridoma Bank), mouse anti-TUJ1 (1:500, 801201, BioLegend), rabbit anti-TUJ1 (1:500, 802001, BioLegend), chicken anti-TUJ1 (1:500, GTX85469, GeneTex), rabbit anti-IBA1 (1:500, 019-19741, FUJIFILM Wako Pure Chemical Corporation), goat anti-IBA1 (1:500, ab5076, abcam), rabbit anti-synaptophysin (1:200, ab14692, abcam), goat anti-ChAT (1:100, ab114P, abcam), rat anti-TREM2 (1:100, MAB17291-100, R&D Systems), rabbit anti-TMEM119 (1:100, ab185337, abcam), rat anti-CD11b (1:100, 101202, BioLegend).

After three washes with PBS-0.1% Triton X-100 for 5min each, coverslips were incubated with corresponding fluorescent secondary antibodies Alexa Fluor 488/568/647 donkey anti-mouse/rabbit/rat/goat, goat anti-chicken (all 1:1000, all ThermoFisher). Coverslips were then washed twice with PBS-0.1% Triton X-100 for 5min each and incubated with 4,6-diamidino-2-phenylindole (DAPI, 1g/mL, Sigma-Aldrich) in PBS for 10min. After an additional 5min-washing step with PBS-0.1% Triton X-100, the coverslips were mounted onto microscopy slides using ProLong Diamond Antifade Mountant (ThermoFisher). Confocal microscopy was then performed using an LSM 710 microscope (Zeiss).

For the analysis of neuronal and MN markers after differentiation, three z-stacks (2m intervals) of randomly selected visual fields (425.1425.1m) were taken for each coverslip at 20magnification. The ratios of TUJ1-positive, ChAT-positive, ISLET1-positive, ChAT-positive/ TUJ1-positive, and ISLET1-positive/ TUJ1-positive cells were then quantified using Fiji in a blinded fashion.

For the analysis of microglial markers in monoculture and co-culture, three z-stacks (1m intervals) of randomly selected visual fields (212.55212.55m) were taken for each coverslip at 40magnification. The expression of CD11b, TMEM119, and TREM2 in IBA1-positive cells in monoculture and co-culture was then quantified using Fiji.

For the analysis of apoptosis in neurons, five z-stacks images of randomly selected visual fields (212.55212.55m) were taken at 40magnification for each coverslip. The ratios of cleaved caspase 3/ TUJ1-positive cells were then quantified for neurons in monoculture and co-culture in a blinded fashion. For the analysis of apoptosis in microglia, three z-stacks images of randomly selected visual fields (212.55212.55m) were taken at 40magnification for each coverslip. The ratios of cleaved caspase 3/ IBA1-positive cells were then quantified for microglia in monoculture and co-culture.

For the analysis of microglial ramifications, five z-stacks images of randomly selected visual fields (212.55212.55m) were taken at 40magnification for each coverslip. To analyze the branching of IBA1-positive microglia in monoculture and co-culture, the average branch length, number of branch points and number of branch endpoints was determined using 3DMorph39, a Matlab-based script for the automated analysis of microglial morphology.

From the same harvest, macrophage precursors (pMacpre) were either lysed directly or differentiated to microglia in monoculture (pMGL) or microglia in co-culture with MNs (co-pMG) for 14days. pMGL were rinsed with PBS and directly lysed in the dish. For both pMacpre and pMGL, RNA was extracted using an RNAeasy Mini Plus kit (Qiagen) according to the manufacturers instructions. Co-cultures were first dissociated by 15min incubation with papain (P4762, Sigma-Aldrich) diluted in accutase (20 U/mL) and gentle trituration based on a previously published protocol40. The cell suspension was then passed through a cell strainer (70m, Falcon) to remove cell clumps. To extract co-pMG, magnetic-activated cell sorting (MACS) was then performed using CD11b-MACS beads (130093-634, Miltenyi Biotec) according to the manufacturers instructions. The panned cell population was lysed for RNA extraction using an RNAeasy Micro kit (Qiagen) according to the manufacturers instructions. In addition, RNA from human fetal microglia and blood monocytes from three different healthy genetic backgrounds wasre-used from our previous study26.

RNA from the four different healthy control lines (listed earlier) per condition (pMacpre, pMGL, co-pMG) was used for RNA sequencing analysis. Material was quantified using RiboGreen (Invitrogen) on the FLUOstar OPTIMA plate reader (BMG Labtech) and the size profile and integrity analysed on the 2200 or 4200 TapeStation (Agilent, RNA ScreenTape). RIN estimates for all samples were between 9.2 and 9.9. Input material was normalised to 100ng prior to library preparation. Polyadenylated transcript enrichment and strand specific library preparation was completed using NEBNext Ultra II mRNA kit (NEB) following manufacturers instructions. Libraries were amplified (14 cycles) on a Tetrad (Bio-Rad) using in-house unique dual indexing primers (based on41). Individual libraries were normalised using Qubit, and the size profile was analysed on the 2200 or 4200 TapeStation. Individual libraries were normalised and pooled together accordingly. The pooled library was diluted to~10nM for storage. The 10nM library was denatured and further diluted prior to loading on the sequencer. Paired end sequencing was performed using a NovaSeq6000 platform (Illumina, NovaSeq 6000 S2/S4 reagent kit, v1.5, 300 cycles), generating a raw read count of a minimum of 34M reads per sample.

Further processing of the raw data was then performed using an in-house pipeline. For comparison, the RNA sequencing data (GSE89189) fromAbud et al.28 and the dataset (GSE85839) fromMuffat et al.29 were downloaded and processed in parallel. Quality control of fastq files was performed using FastQC (https://www.bioinformatics.babraham.ac.uk/projects/fastqc/) and MultiQC42. Paired-end reads were mapped to the human GRCh38.p13 reference genome (https://www.gencodegenes.org) using HISAT2 v2.2.143. Mapping quality control was done using SAMtools44 and Picard (http://broadinstitute.github.io/picard/) metrics. The counts table was obtained using FeatureCounts v2.0.145. Normalization of counts and differential expression analysis for the comparison of pMGL and co-pMG was performed using DESeq2 v1.28.146 in RStudio 1.4.1103, including the biological gender in the model and with the BenjaminiHochberg method for multiple testing correction. Exploratory data analysis was performed following variance-stabilizing transformation of the counts table, using heat maps and hierarchical clustering with the pheatmap 1.0.12 package (https://github.com/raivokolde/pheatmap) and principal component analysis. Log2 fold change (log2 fc) shrinkage for the comparison of pMGL and co-pMG was performed using the ashr package v2.2-4747. Genes with |log2 fc|>2 and adjusted p value<0.01 were defined as differentially expressed and interpreted with annotations from the Gene Ontology database using clusterProfiler v3.16.148 to perform over-representation analyses.

Equal amounts of RNA (30ng) were reverse-transcribed to cDNA using the High-Capacity cDNA Reverse Transcription Kit (ThermoFisher) according to the manufacturers instructions. Quantitative real-time PCR was performed with Fast SYBR Green Master Mix (ThermoFisher) according to the manufacturers instructions using a LightCycler 480 PCR System (Roche). The following primers (ChAT from Eurofins Genomics, all others from ThermoFisher) were used:

Quantification of the relative fold gene expression of samples was performed using the 2Ct method with normalization to the GAPDH reference gene.

AH016-3 Lenti-IP-RFP-microglia were co-cultured with healthy control motor neurons in PEI- and Geltrex-coated glass bottom dishes for confocal microscopy (VWR). The RFP signal was used to identify microglia in co-culture. To visualize microglial movement, images of the RFP signal and brightfield were taken every~30s for 1h (22 stitched images, 20magnification) using a Cell Observer spinning disc confocal microscope (Zeiss) equipped with an incubation system (37C, 5% CO2). To image phagocytic activity, co-cultures were rinsed with Live Cell Imaging Solution (1X, ThermoFisher), and pHrodo Green Zymosan Bioparticles Conjugates (P35365, ThermoFisher) diluted in Live Cell Imaging Solution (50g/mL), which become fluorescent upon phagocytic uptake, were added. The dish was immediately transferred to the spinning disc confocal microscope, and stitched images (33, 20magnification) were acquired every 5min for 2h.

To induce pro-inflammatory (M1) or anti-inflammatory (M2) microglial phenotypes, cells were treated with Lipopolysaccharides (LPS, 100ng/mL, Sigma) and Interferon- (IFN-, 100ng/mL, ThermoFisher), or Interleukin-4 (IL-4, 40ng/mL, R&D Systems) and Interleukin-13 (IL-13, 20ng/mL, Peprotech), respectively, for 18h. Vehicle-treated (co-culture medium) cells were used as an unstimulated (M0) control.

To analyze the clustering of microglia upon pro-inflammatory and anti-inflammatory stimulation, RFP-positive microglia were imaged directly before the addition of M1/M2 inducing agents, and at 9h and 18h post-stimulation using the Cell Observer spinning disc confocal microscope (55 stitched images, 10magnification). The number of individual microglial cells and size of microglial clusters was quantified using the analyze particle function in Fiji.

After stimulation with M1/M2-inducing agents, culture supernatants were collected and spun down at 1200g for 10min at 4C. Pooled samples from three different healthy control lines for each cell type were analyzed using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems) according to the manufacturers instructions. The signal was visualized on a ChemiDoc MP imaging system (Bio-Rad) and analyzed using ImageStudioLite v5.2.5 (LI-COR). Data was then plotted as arbitrary units using the pheatmap 1.0.12 package in RStudio 1.4.1103.

In addition, to confirm the relative expression of Serpin E1 and CHI3L1 in cell culture supernatants, the Human Human Chitinase 3-like 1 Quantikine ELISA Kit (DC3L10) and Human Serpin E1/PAI-1 Quantikine ELISA Kit (DSE100, both R&D Systems) were used according to the manufacturers instructions.

pNeuron, pMGL and co-cultures were plated and maintained in WillCo-dish Glass Bottom Dishes (WillCo Wells) for 14days. Calcium transients were measured using the fluorescent probe Fluo 4-AM according to the manufacturers instructions (ThermoFisher). Cells were incubated with 20M Fluo 4-AM resuspended in 0.2% dimethyl sulfoxide for 30min at RT in Live Imaging Solution (ThermoFisher). After a washing step with Live Imaging Solution, cells were allowed to calibrate at RT for 1520min before imaging. Ca2+ images were taken by fluorescence microscopy at RT. The dye was excited at 488nm and images were taken continuously with a baseline recorded for 30s before stimulation. The stimuli used for calcium release were 50mM KCl (Sigma-Aldrich) for 30s, followed by a washing step for one minute. Microglial calcium release was stimulated by 50M ADP (Merck) under continuous perfusion for 1min, followed by a 1-min wash. Analysis of fluorescence intensity was performed using Fiji. Fluorescence measurements are expressed as a ratio (F/Fo) of the mean change in fluorescence (F) at a pixel relative to the resting fluorescence at that pixel before stimulation (Fo). The responses were analysed in 2040 cells per culture.

MNs on DIV 3345 were maintained in a bath temperature of 25C in a solution containing 167mM NaCl, 2.4mM KCl, 1mM MgCl2, 10mM glucose, 10mM HEPES, and 2mM CaCl2 adjusted to a pH of 7.4 and 300mOsm. Electrodes with tip resistances between 3 and 7M were produced from borosilicate glass (0.86mm inner diameter; 1.5mm outer diameter). The electrode was filled with intracellular solution containing 140mMK-Gluconate, 6mM NaCl, 1mM EGTA, 10mM HEPES, 4mM MgATP, 0.5mM Na3GTP, adjusted to pH 7.3 and 290mOsm. Data acquisition was performed using a Multiclamp 700B amplifier, digidata 1550A and clampEx 6 software (pCLAMP Software suite, Molecular Devices). Data was filtered at 2kHz and digitized at 10kHz. Series resistance (Rs) was continuously monitored and only recordings with stable<50 M and Rs<20% were included in the analysis. Voltage gated channel currents were measured on voltage clamp, neurons were pre-pulsed for 250ms with 140mV and subsequently a 10mV-step voltage was applied from 70 to+70mV. Induced action potentials were recorded on current clamp, neurons were held at 70mV and 8 voltage steps of 10mV, from 10 to 60mV, were applied. Data was analyzed using Clampfit 10.7 (pCLAMP Software suite).

Statistical analyses were conducted using GraphPad Prism 9 (GraphPad Software, San Diego, California USA, http://www.graphpad.com). Comparisons of two groups were performed by two-tailed unpaired t-tests and multiple group comparisons by one-way or two-way analysis of variance (ANOVA) with appropriate post-hoc tests as indicated in the figure legends. The statistical test and number of independent experiments used for each analysis are indicated in each figure legend. Data are presented as single data points and meansSEM. Differences were considered significant when P<0.05 (*P<0.05; **P<0.01; ***P<0.001; ns: not significant). GraphPad Prism 9 or RStudio 1.4.1103 were used to plot data. Final assembly and preparation of all figures was done using Adobe Illustrator 25.4.1.

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Human iPSC co-culture model to investigate the interaction between microglia and motor neurons | Scientific Reports - Nature.com

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Blobs of human brain cells cultivated in the lab, known as brain organoids or mini-brains, are transforming our understanding of neural development and disease. Now, researchers are working to make them more like the real thing

By Clare Wilson

Neil Webb

A DOZEN tiny, creamy balls are suspended in a dish of clear, pink liquid. Seen with the naked eye, they are amorphous blobs. But under a powerful microscope, and with some clever staining, their internal complexity is revealed: intricate whorls and layers of red, blue and green.

These are human brain cells, complete with branching outgrowths that have connected with one other, sparking electrical impulses. This is the stuff that thoughts are made of. And yet, these collections of cells were made in a laboratory in this case, in the lab of Madeline Lancaster at the University of Cambridge.

The structures, known as brain organoids or sometimes mini-brains, hold immense promise for helping us understand the brain. They have already produced fresh insights into how this most mysterious organ functions, how it differs in people with autism and how it goes awry in conditions such as dementia and motor neurone disease. They have even been made to grow primitive eyes.

To truly fulfill the potential of mini-brains, however, neuroscientists want to make them bigger and more complex. Some are attempting to grow them with blood vessels. Others are fusing two organoids, each mimicking a different part of the brain. Should they succeed, their lab-grown brains could model development and disease in the real thing in greater detail than ever before, paving the way to new insights and treatments.

But as researchers seek to make mini-brains genuinely worthy of the name, they move ever closer to a crucial question: at what point will their creations approach sentience?

The key to developing organoids was the discovery of stem cells,

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What lab-grown cerebral organoids are revealing about the brain - New Scientist

Spinal Cord Stem Cells Comments Off on What lab-grown cerebral organoids are revealing about the brain – New Scientist | July 25th, 2022

Glioblastoma (GBM) is a fast-growing type of central nervous system tumour that forms from glial (supportive) tissue of the brain and spinal cord, with cells that look very different from normal ones, said Dr Ata Ul Aleem Bhatti, ex-instructor neurosurgeon, Aga Khan University Medical College, Dar as Salaam, Tanzania, and consultant neurosurgeon at the South City Hospital, Karachi.

Addressing a public awareness seminar on World GBM Day 2022 in collaboration with the Neurospinal & Cancer Care Postgraduate Institute, he said: Like most brain tumors, GBM grow more rapidly than their benign counterparts and affect the brain in many different ways depending on the part of the brain they are located.

Dr Bhatti further explained: Unfortunately, like most cancers in other parts of the body, the exact cause of GBM is unknown. Glioblastoma itself is not the only form of brain cancer, though it is the most common and most aggressive type. Other malignant brain tumours include medulloblastomas, lymphomas and anaplastic astrocytomas, to mention a few.

Various risk factors linked to developing cancer in the brain include over exposure to radiation and some rare inherited conditions. In all of these cases, the exact connection or link remains a mystery, but we do see a pattern of occurrence.

Again, unfortunately, there are no symptoms that will immediately tell someone they are developing a malignant brain tumour, however, there are some common things to look out for, when a person develops a mass or growth in the brain, either benign or malignant. These include a bad headache, but not the type one gets after spending hours in Karachi traffic or a stressful day. This headache is usually worse in the morning and persistent over several weeks. It may be associated with a feeling of wanting to vomit (nausea) or actually vomiting, which tends to make the person feel better.

Unfortunately, according to Dr Bhatti, at the moment there is no cure for brain cancers. While there are many therapies that are being tried and a lot of experimental work going on, we are yet to find a cure.

Malignant brain tumours are usually treated with a combination of surgery, radiotherapy and chemotherapy.

Sometimes, newer options like hormone therapy, immune therapy and others are also used. Which option is offered depends on the type of cancer involved. Surgery remains a main part of any treatment regime for GBM, since it allows for accurate diagnosis and also reduces the amount of tumour the body has to fight against.

In some cases, an attempt is made to remove as much of the tumour as possible to allow the radiotherapy and chemotherapy be more effective.

Dr Adeel Ahmed Memon, consultant clinical & radiation oncologist and assistant professor at the Karachi Institute of Radiotherapy & Nuclear Medicine (KIRAN), gave a radiation oncologist perspective for GBM.

Radiosurgery is a treatment method that uses specialized radiation delivery systems to focus radiation at the site of the tumor, while minimizing the radiation dose to the surrounding brain. Radiosurgery may be used in selective cases for tumor recurrence, often using additional information derived from MRS or PET scans, he said.

Studies have shown that radiation therapy provides most patients with improved outcomes and longer survival rates when given the combination of surgery, radiation and chemotherapy compared with surgery alone. Radiation also may be used as the sole treatment when a glioblastoma tumor is in an area that is not appropriate for surgery.

Guest speaker Dr Reena Kumari, consultant medical oncologist & assistant professor at Dr Ziauddin University Hospital, also shared her views regarding the role of chemotherapy, targeted & immunotherapy and discussed why GBM was difficult to treat brain tumor.

When treating GBM, she explained, what makes treatment challenging is that you have tumor cells that are not active, meaning they are dormant. These cells are known as cancer stem cells and since they are not active they do not die by radiation and chemotherapy.

Unlike other cancers such as breast or lung, brain tumors are extremely genetically heterogeneous means there is a high degree of variation within the same tumor cells that makes each individual glioblastoma molecularly distinct. This can be challenging when predicting prognosis and treatment, if it is in an area which is difficult access, or too close to major blood vessels or other important centers of the brain, it can make surgery tough, tendency of the tumor to come back aggressively is also a great challenge.

A promising targeted treatment is the anti-vascular endothelial growth factor (VEGF) monoclonal antibody bevacizumab. It has been approved by FDA for several different types of cancer, including. Angiogenesis is a key survival feature of many cancers as tumors rely on nutrients from the vasculature to proliferate

A clinical trial has found that selinexor, the first of a new class of anti-cancer drugs called selective inhibitors of nuclear export (SINE) , is able to shrink tumors in almost a third of patients with recurrent glioblastoma,

Dr Kumari urged people to be careful, saying: Negligence in treatment of diseases like GBM can be fatal. She further said that timely treatment of brain tumor was very important as chances of relapsing increases with the grade of tumor.

Dr Sadia Afsar, in-Charge, Neurosurgery Department, Abbasi Shaheed Hospital , highlighted the problems faced by patients with GBM and other brain tumors as this is ignored by community.

Government needs to realise that these conditions are quite common and provide more facilities for early diagnosis and treatment of GBM & other types of brain tumors like MRI, CT-Scan & PET-CT Scanner must be readily available across the country to enhance diagnosis.

The scarcity of Radiotherapy modalities in the country has already been highlighted by her and said that a huge time is wasted in long queue, additionally. The teaching hospitals need to also be equipped to perform proper neurosurgery department and OT, as this is the first step in any treatment programme for brain tumors, including GMBs.

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Negligence in treatment of diseases like glioblastoma can be fatal, seminar told - The News International

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Mesenchymal stem cell isolation and expansion

Bone marrow-derived MSCs were isolated from young (6 weeks) and old (1824 months) C57 black male mice using established techniques42,43 under a protocol approved by the Johns Hopkins University Animal Care and Use Committee. Briefly, immediately following euthanasia, whole bone marrow was flushed out from the bilateral tibias and femurs. After washing by centrifugation at 400g for 10min, cells were plated at 5 106 viable cells per ml. The culture was kept in humidified 5% CO2 incubator at 37C for 72h, when non-adherent cells were removed by changing the media.

All MSC preparations were evaluated using flow cytometry with PE or FITC-conjugated antibodies against murine Sca-1 (1:200; BioLegend 122507), CD31 (1:200; Fisher Scientific BDB554473), CD34 (1:100; eBioscience 14-0341-82), CD44 (1:100; BioLegend 103007), CD45 (1:100; BioLegend 103105), and IgG (1:100; BioLegend 400607) performed on BD LSRII (Becton Dickinson) using DIVA software. At least 10000 events were collected. FlowJo software was used to analyze and create the histograms.

Assessment for osteogenic and adipogenic differentiation was performed using established techniques43. Briefly, to induce osteogenic differentiation, old and young MSCs were seeded into 6-well plates at 1.3 104 cells/well. After 24h the media was replaced with osteogenic differentiation medium containing Iscoves medium supplemented with 100nM dexamethasone, 10mM beta-glycerophosphate, 50 M ascorbic acid, and 1% antibiotic/antimycotic. Cells were maintained in induction media with media changes every 2 days. After 14 days cells fixed in 10% formalin for 15min and calcium deposition was assessed using von Kossa staining. Calcium deposition was then quantified using a colorimetric calcium assay (Calcium CPC Liquicolour Kit StanBio, Boerne, TX) according to the manufacturers instructions. To induce adipogenic differentiation, old and young MSCs were seeded in 6-well plates at 2 105 cells/well. When confluent the media was replaced with adipogenic induction medium containing DMEM-HG, 10% FBS, 5% rabbit serum, 1uM dexamethasone, 10g/mL insulin, 200 M indomethacin, 500 M isobutylmethylxanthine (IBMX), and antibiotic/antimycotic for 3 days followed by exposure to followed by exposure to adipogenic maintenance medium (DMEM-HG, 10% FBS, insulin 10g/ml and P/S) for 3 days. After 3 cycles of induction and maintenance exposure cells were rinsed with PBS and fixed in 10% formalin for 10min. The cells were then stained with Oil Red O to assess for lipid droplets. After imaging Oil Red O extraction was performed using 100% isopropanol. Extract samples were transferred to a 96-well plate and absorbance readings were taken at 490nm to quantify extracted Oil Red O.

Confirmed MSCs were expanded in culture in media prepared by combining 490ml Medium 200 PRF (Gibco Invitrogen, Carlsbad, CA), a standard basal medium intended for culture of large vessel human endothelial cells, with 10ml Low Serum Growth Supplement (LSGS; Gibco Invitrogen). The final preparation contained 2% fetal bovine serum (FBS), 3ng/ml basic fibroblast growth factor (bFGF), 10ng/ml human epidermal growth factor, 10g/ml heparin, and 1g/ml hydrocortisone. Cells were incubated under standard conditions (5% CO2 and 37C). Expanded MSCs at low passage numbers (P2-P5) were used for the experiments. In the event frozen cells were used, they were thawed and grown for one passage prior to use in the experiments.

To prevent cell-cell interaction and assess only paracrine-mediated effects (i.e. those resulting from release of soluble factors), angiogenesis experiments were performed using bioreactor tubes (BT) constructed with CellMax semi-permeable polysulfone membrane tubing (Spectrum Labs, Rancho Dominguez, CA). These allowed the free diffusion of soluble proteins and other molecules released by the cells up to a 500kDa molecular weight cut-off, but not of the cells themselves. To load BTs, MSCs were trypsinized and suspended in Medium 200 PRF without LSGS supplementation (i.e. media devoid of stimulatory growth factors). MSCs were counted using a Scepter automated cell counter (Millipore, Billerica, MA), which had been previously standardized for accuracy. The desired number of MSCs was spun down and resuspended to a total volume of 100 ul that was injected into the BTs using a 0.5mL syringe. To compare paracrine-mediated angiogenesis by old and young MSCs, BTs were loaded with either 105 old or 105 young MSCs. Once cell injection was complete, the tubes were heat-sealed at both ends and the MSC-loaded tubes, fully submerged in media, were grown at standard culture conditions (37C, 5% CO2) for 7 days (Fig. 3a).

ELISA assays were performed to measure paracrine factor (PF) production by the MSCs contained within the BTs grown in culture. Tubes loaded with 2 105 MSCs were submerged in 5mL of alpha-MEM basal medium (Stemcell Technologies, Tukwila, WA) supplemented with 20% FBS (Gibco Invitrogen, Carlsbad, CA) in a 6-well plate. At day 7, conditioned media was collected from each well, spun down for 1min to pellet any debris, and then flash frozen at 80C. Conditioned media samples were assessed for the concentrations of vascular endothelial growth factor (VEGF), stromal derived factor-1 (SDF1) and insulin-like growth factor-1 (IGF1) by ELISA (Quantikine, R&D Systems, Minneapolis, MN) according to the manufacturers instructions.

BTs were removed at day 7 and placed in separate wells of a 6-well plate containing human umbilical vein endothelial cells (HUVECs)44. Briefly, 105 HUVECs (Gibco Invitrogen, Carlsbad, CA) suspended in Medium 200PRF were plated per well in Geltrex (Gibco Invitrogen) coated 6-well plates. Negative control wells received a bioreactor loaded with un-supplemented Medium 200PRF only (i.e. no cells). Positive control wells were plated with 105 HUVECs suspended in 1mL of Medium 200PRF supplemented with LSGS, which is known to induce HUVEC tubule formation. After 18h at standard culture conditions (37C, 5% CO2), the wells were imaged to allow quantitative analysis of the resultant HUVEC tubule network. Images were taken in the center of each well and in all four quadrants at pre-determined locations (5 pictures total), at 100x magnification. The total length of the tubule networks captured in the images of each well was measured using ImageJ software. To allow for comparisons between experiments, the total length of the tubule network in each well was normalized to the average length of the tubule network in the negative control wells, and reported as a normalized ratio.

To assess the effect of young MSC-generated PFs on PF-mediated angiogenesis by old MSCs, BTs were prepared as described above containing either 105 young or 105 old MSCs. Two BTs were placed together in a 6-well plate in 5mL MSC media and incubated for 7 days at standard culture conditions (Fig. 3b) using a BT containing old MSCs paired with either a separate BT with other old MSCs (control) or a separate BT with young MSCs. After 7 days the tubes were removed, washed with un-supplemented Medium 200 PRF, and then used separately in the HUVEC assay as described above. After the HUVEC assay was complete (18h) the BTs were placed in separate wells of 6-well plates and grown in culture for 7 additional days with collection of conditioned media for PF release.

Replicates of 105 old MSCs were cultured separately, or in co-culture with young MSCs, for 7 days using a 0.4m Transwell system in 6-well plates (Corning), which allow transfer of soluble paracrine factors released by the cells, but not of the cells themselves. Following RNA purification, library preparation, amplification, and Illumina sequencing, the open source Galaxy pipeline was used for data processing and analyses. After alignment of raw sequencing reads to the UCSC mm10 genome using HISAT2, transcript assembly, alignment quantification, count normalization, and differential expression analyses were conducted with StringTie, featureCounts, DESeq2, and Genesis. Quantitative PCR (KAPA SYBR FAST One-Step qRT-PCR, Wilmington, MA) was used to validate 24 transcripts identified by RNA sequencing. Target genes were selected based on their presence in significantly regulated pathways and quantified relative to 18S ribosomal RNA using the 2Ct method45.

To validate the results of the RNA sequencing and RT-PCR results, a functional autophagy assay was performed to assess relative autophagy between old, young, and rejuvenated old MSCs. Old, young and rejuvenated cells were cultured (or co-cultured, in the case of rejuvenated cells) for 7 days in 6-well plates (105 cells per well). On Day 8, cells were trypsinzed, counted and 104 cells were transferred to each well of a 96-well black plate with clear bottom and incubated for 6h. The Autophagy Assay Kit (Sigma Aldrich, St. Louis, MO) measures autophagy using a proprietary fluorescent autophagosome marker in a microplate reader (ex=360; em=520nm). Three separate experiments were performed in triplicate each for each condition. To account for possible differences introduced by counting cells, results for each cell type were normalized based on absorbance (450nm) of a Cell Counting Kit-8 (Dojindo Molecular Technologies, Inc. Rockville, MD).

Data are reported as mean standard error of the mean (SEM) unless otherwise indicated. Comparisons between groups for the HUVEC experiments were performed using the permutation test. For the PF ELISA data, groups were compared using the MannWhitney test. The autophagy assay and rt-PCR results were assessed using two-tailed t tests. For these experiments a p-value < 0.05 was deemed significant. In the RNA sequencing differential expression analysis, a false discovery rate (FDR) of <0.05 was considered significant.

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Paracrine-mediated rejuvenation of aged mesenchymal stem cells is associated with downregulation of the autophagy-lysosomal pathway | npj Aging -...

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In short, stem cells initiate the production of new tissue cells, which can then replace their diseased counterparts.

Mesenchymal stem cells (MSCs) are adult stem cells found in many areas of the body such as bone marrow. The unique thing about these cells is their compatibility with a range of tissues such as bone, cartilage, muscle, or fat. MSCs respond to injury or disease by migrating to these damaged areas, where they restore tissue function by replacing the damaged cells.

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It has recently been shown that the success of MSCs relies on their ability to release cell signals their mechanism to initiate tissue regeneration. These signals are packaged into extracellular vehicles (EVs) which are essentially bubbles of information. These are released by MSCs and taken up by the injured or diseased tissue cells to kickstart their inbuilt process of regeneration.

Through funding from the Royal Society of Edinburgh, research has started into the development of artificial EVs as a viable alternative to cell therapy. These EVs will contain the key molecules released by stem cells when they are responding to injury cues in the body.

The power to induce tissue regeneration would provide a significant new tool in biomedical treatment, such as incorporating EVs into synthetic hydrogels within a wound dressing to encourage and accelerate healing.

Within the lab setting, we have been able to manipulate stem cell cultures to produce EVs with different signal make-ups, and accurately identify their properties.

Controlling and identifying the different make-ups contained in EV signals which in turn induce different cell responses is crucial if we want to operationalise their use in medicine.

We now aim to synthesise artificial vesicles, or bubbles, for different clinical problems, such as, for example, bubbles with potent wound-healing properties that would help our ability to use new artificial stem cell therapy.

The research is underway and it is showing promise that we may be able to harness the regenerative power of stem cells in the near future.

An artificial EV-based approach also has several advantages over stem cell-based therapies, such as having increased potency and greater consistency in treatment, and at a lower cost to carry out.

Both inside and on the surface of the body, we would have the ability to induce a process vital to medical treatment we work with every day and, in turn, open a whole new avenue of possibilities in biomedical science.

Dr Catherine Berry is a reader in the Centre for the Cellular Microenvironment at the University of Glasgow, and a recipient of the Royal Society of Edinburghs personal research fellowship in 2021. This article expresses her own views. The RSE is Scotland's national academy, bringing great minds together to contribute to the social, cultural and economic well-being of Scotland. Find out more at rse.org.uk and @RoyalSocEd.

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Stem cells: Could we gain the power to induce cell regeneration? Dr Catherine Berry - The Scotsman

Bone Marrow Stem Cells Comments Off on Stem cells: Could we gain the power to induce cell regeneration? Dr Catherine Berry – The Scotsman | July 25th, 2022

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United States of AmericaUS Virgin IslandsUnited States Minor Outlying IslandsCanadaMexico, United Mexican StatesBahamas, Commonwealth of theCuba, Republic ofDominican RepublicHaiti, Republic ofJamaicaAfghanistanAlbania, People's Socialist Republic ofAlgeria, People's Democratic Republic ofAmerican SamoaAndorra, Principality ofAngola, Republic ofAnguillaAntarctica (the territory South of 60 deg S)Antigua and BarbudaArgentina, Argentine RepublicArmeniaArubaAustralia, Commonwealth ofAustria, Republic ofAzerbaijan, Republic ofBahrain, Kingdom ofBangladesh, People's Republic ofBarbadosBelarusBelgium, Kingdom ofBelizeBenin, People's Republic ofBermudaBhutan, Kingdom ofBolivia, Republic ofBosnia and HerzegovinaBotswana, Republic ofBouvet Island (Bouvetoya)Brazil, Federative Republic ofBritish Indian Ocean Territory (Chagos Archipelago)British Virgin IslandsBrunei DarussalamBulgaria, People's Republic ofBurkina FasoBurundi, Republic ofCambodia, Kingdom ofCameroon, United Republic ofCape Verde, Republic ofCayman IslandsCentral African RepublicChad, Republic ofChile, Republic ofChina, People's Republic ofChristmas IslandCocos (Keeling) IslandsColombia, Republic ofComoros, Union of theCongo, Democratic Republic ofCongo, People's Republic ofCook IslandsCosta Rica, Republic ofCote D'Ivoire, Ivory Coast, Republic of theCyprus, Republic ofCzech RepublicDenmark, Kingdom ofDjibouti, Republic ofDominica, Commonwealth ofEcuador, Republic ofEgypt, Arab Republic ofEl Salvador, Republic ofEquatorial Guinea, Republic ofEritreaEstoniaEthiopiaFaeroe IslandsFalkland Islands (Malvinas)Fiji, Republic of the Fiji IslandsFinland, Republic ofFrance, French RepublicFrench GuianaFrench PolynesiaFrench Southern TerritoriesGabon, Gabonese RepublicGambia, Republic of theGeorgiaGermanyGhana, Republic ofGibraltarGreece, Hellenic RepublicGreenlandGrenadaGuadaloupeGuamGuatemala, Republic ofGuinea, RevolutionaryPeople's Rep'c ofGuinea-Bissau, Republic ofGuyana, Republic ofHeard and McDonald IslandsHoly See (Vatican City State)Honduras, Republic ofHong Kong, Special Administrative Region of ChinaHrvatska (Croatia)Hungary, Hungarian People's RepublicIceland, Republic ofIndia, Republic ofIndonesia, Republic ofIran, Islamic Republic ofIraq, Republic ofIrelandIsrael, State ofItaly, Italian RepublicJapanJordan, Hashemite Kingdom ofKazakhstan, Republic ofKenya, Republic ofKiribati, Republic ofKorea, Democratic People's Republic ofKorea, Republic ofKuwait, State ofKyrgyz RepublicLao People's Democratic RepublicLatviaLebanon, Lebanese RepublicLesotho, Kingdom ofLiberia, Republic ofLibyan Arab JamahiriyaLiechtenstein, Principality ofLithuaniaLuxembourg, Grand Duchy ofMacao, Special Administrative Region of ChinaMacedonia, the former Yugoslav Republic ofMadagascar, Republic ofMalawi, Republic ofMalaysiaMaldives, Republic ofMali, Republic ofMalta, Republic ofMarshall IslandsMartiniqueMauritania, Islamic Republic ofMauritiusMayotteMicronesia, Federated States ofMoldova, Republic ofMonaco, Principality ofMongolia, Mongolian People's RepublicMontserratMorocco, Kingdom ofMozambique, People's Republic ofMyanmarNamibiaNauru, Republic ofNepal, Kingdom ofNetherlands AntillesNetherlands, Kingdom of theNew CaledoniaNew ZealandNicaragua, Republic ofNiger, Republic of theNigeria, Federal Republic ofNiue, Republic ofNorfolk IslandNorthern Mariana IslandsNorway, Kingdom ofOman, Sultanate ofPakistan, Islamic Republic ofPalauPalestinian Territory, OccupiedPanama, Republic ofPapua New GuineaParaguay, Republic ofPeru, Republic ofPhilippines, Republic of thePitcairn IslandPoland, Polish People's RepublicPortugal, Portuguese RepublicPuerto RicoQatar, State ofReunionRomania, Socialist Republic ofRussian FederationRwanda, Rwandese RepublicSamoa, Independent State ofSan Marino, Republic ofSao Tome and Principe, Democratic Republic ofSaudi Arabia, Kingdom ofSenegal, Republic ofSerbia and MontenegroSeychelles, Republic ofSierra Leone, Republic ofSingapore, Republic ofSlovakia (Slovak Republic)SloveniaSolomon IslandsSomalia, Somali RepublicSouth Africa, Republic ofSouth Georgia and the South Sandwich IslandsSpain, Spanish StateSri Lanka, Democratic Socialist Republic ofSt. HelenaSt. Kitts and NevisSt. LuciaSt. Pierre and MiquelonSt. Vincent and the GrenadinesSudan, Democratic Republic of theSuriname, Republic ofSvalbard & Jan Mayen IslandsSwaziland, Kingdom ofSweden, Kingdom ofSwitzerland, Swiss ConfederationSyrian Arab RepublicTaiwan, Province of ChinaTajikistanTanzania, United Republic ofThailand, Kingdom ofTimor-Leste, Democratic Republic ofTogo, Togolese RepublicTokelau (Tokelau Islands)Tonga, Kingdom ofTrinidad and Tobago, Republic ofTunisia, Republic ofTurkey, Republic ofTurkmenistanTurks and Caicos IslandsTuvaluUganda, Republic ofUkraineUnited Arab EmiratesUnited Kingdom of Great Britain & N. IrelandUruguay, Eastern Republic ofUzbekistanVanuatuVenezuela, Bolivarian Republic ofViet Nam, Socialist Republic ofWallis and Futuna IslandsWestern SaharaYemenZambia, Republic ofZimbabwe

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He's the match: Arconic employee gets call 20 years after signing up to be bone marrow donor - Maryville Daily Times

Bone Marrow Stem Cells Comments Off on He’s the match: Arconic employee gets call 20 years after signing up to be bone marrow donor – Maryville Daily Times | July 25th, 2022

The new report by Expert Market Research titled, Global Stem Cell Banking Market Report and Forecast 2021-2026, gives an in-depth analysis of the globalstem cell banking market, assessing the market based on its segments like Service type, product type, utilisation, bank type, application, and major regions like Asia Pacific, Europe, North America, Middle East and Africa and Latin America. The report tracks the latest trends in the industry and studies their impact on the overall market. It also assesses the market dynamics, covering the key demand and price indicators, along with analysing the market based on the SWOT and Porters Five Forces models.

Request a free sample copy in PDF or view the report summary@https://bityl.co/CPix

The key highlights of the report include:

Market Overview (2021-2026)

The global stem cell bank market is primarily driven by the advancements in the field of medicine and the rising prevalence of genetic and degenerativediseases. Further, the increasing research and development of more effective technologies for better preservation, processing, and storage of stem cells are aiding the growth. Additionally, rising prevalence of chronic diseases globally is increasing the for advances inmedicaltechnologies, thus pushing the growth further. Moreover, factors such as rising health awareness, developinghealthcare infrastructure, growing geriatric population, and the inflatingdisposableincomes are expected to propel the market in the forecast period.

Industry Definition and Major Segments

Stem cells are undifferentiated cells present in bone marrow,umbilical cordadipose tissue and blood. They have the ability to of differentiate and regenerate. The process of storing and preserving these cells for various application such as gene therapy, regenerative medicine and tissue engineering is known as stem cell banking.

Explore the full report with the table of contents@https://bityl.co/CPiy

By service type, the market is divided into:

Based on product type, the industry can be segmented into:

The market is bifurcated based on utilization into:

By bank type, the industry can be broadly categorized into:

Based on application, the industry can be segmented into:

On the basis of regional markets, the industry is divided into:

1 North America1.1 United States of America1.2 Canada2 Europe2.1 Germany2.2 United Kingdom2.3 France2.4 Italy2.5 Others3 Asia Pacific3.1 China3.2 Japan3.3 India3.4 ASEAN3.5 Others4 Latin America4.1 Brazil4.2 Argentina4.3 Mexico4.4 Others5 Middle East & Africa5.1 Saudi Arabia5.2 United Arab Emirates5.3 Nigeria5.4 South Africa5.5 Others

Market Trends

Regionally, North America is projected to dominate the global stem cell bank market and expand at a significant rate. This can be attributed to increasing research and development for stem cell application in various medical fields. Further, growing investments of pharmaceutical players and development infrastructure are other factors that are expected to stem cell bank market in the region. Meanwhile, Asia Pacific market is also expected to witness fast growth owing to the rapid development in healthcare facilities and increasing awareness of stem cell banking in countries such as China, India, and Indonesia.

Key Market Players

The major players in the market are Cryo-Cell International, Inc., Smart Cells International Ltd., CSG-BIO Company, Inc., CBR Systems Inc., ViaCord, LLC, LifeCell International Pvt. Ltd., and a few others. The report covers the market shares, capacities, plant turnarounds, expansions, investments and mergers and acquisitions, among other latest developments of these market players.

About Us:

Expert Market Research (EMR) is leading market research company with clients across the globe. Through comprehensive data collection and skilful analysis and interpretation of data, the company offers its clients extensive, latest and actionable market intelligence which enables them to make informed and intelligent decisions and strengthen their position in the market. The clientele ranges from Fortune 1000 companies to small and medium scale enterprises.

EMR customises syndicated reports according to clients requirements and expectations. The company is active across over 15 prominent industry domains, including food and beverages, chemicals and materials, technology and media, consumer goods, packaging, agriculture, and pharmaceuticals, among others.

Over 3000 EMR consultants and more than 100 analysts work very hard to ensure that clients get only the most updated, relevant, accurate and actionable industry intelligence so that they may formulate informed, effective and intelligent business strategies and ensure their leadership in the market.

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*We at Expert Market Research always thrive to give you the latest information. The numbers in the article are only indicative and may be different from the actual report.

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Global Stem Cell Banking Market To Be Driven At A CAGR Of 13.5% In The Forecast Period Of 2021-2026 This Is Ardee - This Is Ardee

Bone Marrow Stem Cells Comments Off on Global Stem Cell Banking Market To Be Driven At A CAGR Of 13.5% In The Forecast Period Of 2021-2026 This Is Ardee – This Is Ardee | July 25th, 2022

SINGAPORE - A Singaporean doctorwho is one of the top cell therapy experts in the worldhas been appointed to a World Health Organisation (WHO) expert panel.

Dr Mickey Koh is so sought-after in his field that for the past 15 years, he has been holding two jobs in two different countries.

The 56-year-old shuttles between England and Singapore, spending six weeks at a time in London, where he oversees the haematology department and looks after bone marrow transplant patients at St George's University Hospital, before returning to Singapore for a week and a half to head the cell therapy programme at the Health Sciences Authority.

Cell therapy is a growing field of medicine that uses living cells as treatment for a variety of diseases and conditions. This is an increasingly important therapeutic area and both his employers have agreed to his unusual schedule.

Over in London, Dr Koh is head of the Haematology Department at St George's Hospital and Medical School. In Singapore, he is the programme and medical director of the cell and gene therapy facility at the Health Sciences Authority.

In May, Dr Koh was selected to be on the WHO Expert Advisory Panel on Biological Standardisation.

Individuals on the panel have to be invited by WHO to apply, and are well recognised in their respective scientific fields. Eminent names on the panel include the current president of the Paul-Ehrlich-Institut in Germany, which is the country's federal agency, medical regulatory body and research institution for vaccines and biomedicine.

The WHO panel, which is made up of about 25 members, provides detailed recommendations and guidelines for the manufacturing, licensing and standardisation of biological products, which include blood, monoclonal antibodies, vaccines and, increasingly, cell-based therapeutics.

The recommendations and advice are passed on to the executive board of the World Health Assembly, which is the decision-making body of WHO.

Dr Koh's role had to be endorsed by the British government and was a direct appointment by the director-general of WHO.

His appointment as a panel expert will last for a term of four years.

Speaking to The Straits Times, Dr Koh shared his thoughts about the importance of regulation: "We are well aware that there is a very lucrative worldwide market peddling unproven stem cell treatments, where side effects are often unknown, and such unregulated practice can result in serious harm.

"This is already happening. People are claiming that you can use stem cells to treat things like ageing, and even very serious conditions like strokes, without any evidence."

With many medications now taking the form of biologics - a drug product derived from biological sources such as cells - the next wave of treatment would be the utilisation of these cells for the treatment of a wide range of diseases, Dr Koh said.

Excerpt from:
S'porean doctor, a sought-after top expert in cell therapy, appointed to WHO expert panel - The Straits Times

Bone Marrow Stem Cells Comments Off on S’porean doctor, a sought-after top expert in cell therapy, appointed to WHO expert panel – The Straits Times | July 25th, 2022

Market Overview

Thecell culture media marketis expected to cross USD 4.33 billion by 2027 at a CAGR of8.33%.

Market Dynamics

The markets growth is being fueled by a diverse range of cell culture media applications, increased research and development in the pharmaceutical industry, an increase in the prevalence of chronic diseases, and increased expansion and product launches by major players. Over the last few decades, advancements in cell culture technology have accelerated. It is widely regarded as one of the most dependable, robust, and mature technologies for biotherapeutic product development.

The high cost of cell culture media and the risk of contamination, on the other hand, are impeding the markets growth. However, the growing emphasis on regenerative and personalized medicine is likely to spur growth in the global cell culture media market.

Get Sample Report: https://www.marketresearchfuture.com/sample_request/4462

Competitive Dynamics

The notable players are the Merck KGaA (Germany), Bio-Rad Laboratories, Inc. (US), Thermo Fisher Scientific Inc. (US), Lonza (Switzerland), GE Healthcare (US), Becton, Dickinson and Company (US), HiMedia Laboratories (India), Corning Incorporated (US), PromoCell (Germany), Sera Scandia A/S (Denmark), The Sartorius Group (Germany), and Fujifilm Holdings Corporation (Japan).

Segmental Analysis

The global market for cell culture media has been segmented according to product type, application, and end user.

The market has been segmented by product type into classical media, stem cell media, serum-free media, and others.

Further subcategories of stem cell culture media include bone marrow, embryonic stem cells, mesenchymal stem cells, and neural stem cells.

The market is segmented into four application segments: drug discovery and development, cancer research, genetic engineering, and tissue engineering and biochemistry.

The market is segmented by end user into biochemistry and pharmaceutical companies, research laboratories, academic institutions, and pathology laboratories.

Regional Overview

According to region, the global cell culture media market is segmented into the Americas, Europe, Asia-Pacific, and the Middle East & Africa.

The Americas dominated the global cell culture media market. The large share is attributed to the presence of major manufacturers, rising disease prevalence resulting in increased demand for drugs and other medications, technological advancements in the preclinical and clinical segments, growing public awareness, and high disposable income.

Europe ranks second in terms of market size for cell culture media. Factors such as an increase in the biopharmaceutical sector in the European region, increased government initiatives to promote research to find a cure for the growing number of chronic diseases, an increase in the number of pharmaceutical manufacturers, improving economies, a high disposable income per individual, and increased healthcare spending are all contributing to the markets growth in this region. The European market is expected to be driven by expanding R&D activities and a developing biopharmaceutical sector.

Asia-Pacific held the third-largest market share, owing to the presence of numerous research organizations, low manufacturing costs, low labor costs, developing healthcare infrastructure, and increased investment by American and European market giants in Asian countries such as China and India.

The Middle East and Africa, with limited economic development and extremely low income, held the smallest market share in 2019 but is expected to grow due to growing public awareness and demand for improved healthcare facilities in countries, as well as rising disposable income.

Browse Full Reports: https://www.marketresearchfuture.com/reports/cell-culture-media-market-4462

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Cell Culture Media Market: Competitive Approach, Breakdown And Forecast by 2027 - Digital Journal

Bone Marrow Stem Cells Comments Off on Cell Culture Media Market: Competitive Approach, Breakdown And Forecast by 2027 – Digital Journal | July 25th, 2022

Tecartus (Brexucabtagene Autoleucel) First and Only CAR T in Europe to Receive Positive CHMP Opinion to Treat Adults 26+ with r/r ALL

If Approved, it will Address a Significant Unmet Need for a Patient Population with Limited Treatment Options

SANTA MONICA, Calif.--(BUSINESS WIRE)--Kite, a Gilead Company (Nasdaq: GILD), today announces that the European Medicines Agency (EMA) Committee for Medicinal Products for Human Use (CHMP) has issued a positive opinion for Tecartus (brexucabtagene autoleucel) for the treatment of adult patients 26 years of age and above with relapsed or refractory (r/r) B-cell precursor acute lymphoblastic leukemia (ALL). If approved, Tecartus will be the first and only Chimeric Antigen Receptor (CAR) T-cell therapy for this population of patients who have limited treatment options. Half of adults with ALL will relapse, and median overall survival (OS) for this group is only approximately eight months with current standard-of-care treatments.

Kites goal is clear: to bring the hope of survival to more patients with cancer around the world through cell therapy, said Christi Shaw, CEO, Kite. Todays CHMP positive opinion in adult ALL brings us a step closer to delivering on the promise that cell therapies have to transform the way cancer is treated.

Following this positive opinion, the European Commission will now review the CHMP opinion; the final decision on the Marketing Authorization is expected in the coming months.

Adults with relapsed or refractory ALL often undergo multiple treatments including chemotherapy, targeted therapy and stem cell transplant, creating a significant burden on a patients quality of life, said Max S. Topp, MD, professor and head of Hematology, University Hospital of Wuerzburg, Germany. If approved, patients in Europe will have a meaningful advancement in treatment. Tecartus has demonstrated durable responses, suggesting the potential for long-term remission and a new approach to care.

Results from the ZUMA-3 international multicenter, single-arm, open-label, registrational Phase 1/2 study of adult patients (18 years old) with relapsed or refractory ALL, demonstrated that 71% of the evaluable patients (n=55) achieved complete remission (CR) or CR with incomplete hematological recovery (CRi) with a median follow-up of 26.8 months. In an extended data set of all patients dosed with the pivotal dose (n=78) the median overall survival for all patients was more than two years (25.4 months) and almost four years (47 months) for responders (patients who achieved CR or CRi). Among efficacy-evaluable patients, median duration of remission (DOR) was 18.6 months. Among the patients treated with Tecartus at the target dose (n=100), Grade 3 or higher cytokine release syndrome (CRS) and neurologic events occurred in 25% and 32% of patients, respectively, and were generally well-managed.

About ZUMA-3

ZUMA-3 is an ongoing international multicenter (US, Canada, EU), single arm, open label, registrational Phase 1/2 study of Tecartus in adult patients (18 years old) with ALL whose disease is refractory to or has relapsed following standard systemic therapy or hematopoietic stem cell transplantation. The primary endpoint is the rate of overall complete remission or complete remission with incomplete hematological recovery by central assessment. Duration of remission and relapse-free survival, overall survival, minimal residual disease (MRD) negativity rate, and allo-SCT rate were assessed as secondary endpoints.

About Acute Lymphoblastic Leukemia

ALL is an aggressive type of blood cancer that develops when abnormal white blood cells accumulate in the bone marrow until there isnt any room left for blood cells to form. In some cases, these abnormal cells invade healthy organs and can also involve the lymph nodes, spleen, liver, central nervous system and other organs. The most common form is B cell precursor ALL. Globally, approximately 64,000 people are diagnosed with ALL each year, including around 3,300 people in Europe.

About Tecartus

Please see full FDA Prescribing Information, including BOXED WARNING and Medication Guide.

Tecartus is a CD19-directed genetically modified autologous T cell immunotherapy indicated for the treatment of:

This indication is approved under accelerated approval based on overall response rate and durability of response. Continued approval for this indication may be contingent upon verification and description of clinical benefit in a confirmatory trial.

U.S. IMPORTANT SAFETY INFORMATION

BOXED WARNING: CYTOKINE RELEASE SYNDROME and NEUROLOGIC TOXICITIES

Cytokine Release Syndrome (CRS), including life-threatening reactions, occurred following treatment with Tecartus. In ZUMA-2, CRS occurred in 91% (75/82) of patients receiving Tecartus, including Grade 3 CRS in 18% of patients. Among the patients who died after receiving Tecartus, one had a fatal CRS event. The median time to onset of CRS was three days (range: 1 to 13 days) and the median duration of CRS was ten days (range: 1 to 50 days). Among patients with CRS, the key manifestations (>10%) were similar in MCL and ALL and included fever (93%), hypotension (62%), tachycardia (59%), chills (32%), hypoxia (31%), headache (21%), fatigue (20%), and nausea (13%). Serious events associated with CRS included hypotension, fever, hypoxia, tachycardia, and dyspnea.

Ensure that a minimum of two doses of tocilizumab are available for each patient prior to infusion of Tecartus. Following infusion, monitor patients for signs and symptoms of CRS daily for at least seven days for patients with MCL and at least 14 days for patients with ALL at the certified healthcare facility, and for four weeks thereafter. Counsel patients to seek immediate medical attention should signs or symptoms of CRS occur at any time. At the first sign of CRS, institute treatment with supportive care, tocilizumab, or tocilizumab and corticosteroids as indicated.

Neurologic Events, including those that were fatal or life-threatening, occurred following treatment with Tecartus. Neurologic events occurred in 81% (66/82) of patients with MCL, including Grade 3 in 37% of patients. The median time to onset for neurologic events was six days (range: 1 to 32 days) with a median duration of 21 days (range: 2 to 454 days) in patients with MCL. Neurologic events occurred in 87% (68/78) of patients with ALL, including Grade 3 in 35% of patients. The median time to onset for neurologic events was seven days (range: 1 to 51 days) with a median duration of 15 days (range: 1 to 397 days) in patients with ALL. For patients with MCL, 54 (66%) patients experienced CRS before the onset of neurological events. Five (6%) patients did not experience CRS with neurologic events and eight patients (10%) developed neurological events after the resolution of CRS. Neurologic events resolved for 119 out of 134 (89%) patients treated with Tecartus. Nine patients (three patients with MCL and six patients with ALL) had ongoing neurologic events at the time of death. For patients with ALL, neurologic events occurred before, during, and after CRS in 4 (5%), 57 (73%), and 8 (10%) of patients; respectively. Three patients (4%) had neurologic events without CRS. The onset of neurologic events can be concurrent with CRS, following resolution of CRS or in the absence of CRS.

The most common neurologic events (>10%) were similar in MCL and ALL and included encephalopathy (57%), headache (37%), tremor (34%), confusional state (26%), aphasia (23%), delirium (17%), dizziness (15%), anxiety (14%), and agitation (12%). Serious events including encephalopathy, aphasia, confusional state, and seizures occurred after treatment with Tecartus.

Monitor patients daily for at least seven days for patients with MCL and at least 14 days for patients with ALL at the certified healthcare facility and for four weeks following infusion for signs and symptoms of neurologic toxicities and treat promptly.

REMS Program: Because of the risk of CRS and neurologic toxicities, Tecartus is available only through a restricted program under a Risk Evaluation and Mitigation Strategy (REMS) called the Yescarta and Tecartus REMS Program which requires that:

Hypersensitivity Reactions: Serious hypersensitivity reactions, including anaphylaxis, may occur due to dimethyl sulfoxide (DMSO) or residual gentamicin in Tecartus.

Severe Infections: Severe or life-threatening infections occurred in patients after Tecartus infusion. Infections (all grades) occurred in 56% (46/82) of patients with MCL and 44% (34/78) of patients with ALL. Grade 3 or higher infections, including bacterial, viral, and fungal infections, occurred in 30% of patients with ALL and MCL. Tecartus should not be administered to patients with clinically significant active systemic infections. Monitor patients for signs and symptoms of infection before and after Tecartus infusion and treat appropriately. Administer prophylactic antimicrobials according to local guidelines.

Febrile neutropenia was observed in 6% of patients with MCL and 35% of patients with ALL after Tecartus infusion and may be concurrent with CRS. The febrile neutropenia in 27 (35%) of patients with ALL includes events of febrile neutropenia (11 (14%)) plus the concurrent events of fever and neutropenia (16 (21%)). In the event of febrile neutropenia, evaluate for infection and manage with broad spectrum antibiotics, fluids, and other supportive care as medically indicated.

In immunosuppressed patients, life-threatening and fatal opportunistic infections have been reported. The possibility of rare infectious etiologies (e.g., fungal and viral infections such as HHV-6 and progressive multifocal leukoencephalopathy) should be considered in patients with neurologic events and appropriate diagnostic evaluations should be performed.

Hepatitis B virus (HBV) reactivation, in some cases resulting in fulminant hepatitis, hepatic failure, and death, can occur in patients treated with drugs directed against B cells. Perform screening for HBV, HCV, and HIV in accordance with clinical guidelines before collection of cells for manufacturing.

Prolonged Cytopenias: Patients may exhibit cytopenias for several weeks following lymphodepleting chemotherapy and Tecartus infusion. In patients with MCL, Grade 3 or higher cytopenias not resolved by Day 30 following Tecartus infusion occurred in 55% (45/82) of patients and included thrombocytopenia (38%), neutropenia (37%), and anemia (17%). In patients with ALL who were responders to Tecartus treatment, Grade 3 or higher cytopenias not resolved by Day 30 following Tecartus infusion occurred in 20% (7/35) of the patients and included neutropenia (12%) and thrombocytopenia (12%); Grade 3 or higher cytopenias not resolved by Day 60 following Tecartus infusion occurred in 11% (4/35) of the patients and included neutropenia (9%) and thrombocytopenia (6%). Monitor blood counts after Tecartus infusion.

Hypogammaglobulinemia: B cell aplasia and hypogammaglobulinemia can occur in patients receiving treatment with Tecartus. Hypogammaglobulinemia was reported in 16% (13/82) of patients with MCL and 9% (7/78) of patients with ALL. Monitor immunoglobulin levels after treatment with Tecartus and manage using infection precautions, antibiotic prophylaxis, and immunoglobulin replacement.

The safety of immunization with live viral vaccines during or following Tecartus treatment has not been studied. Vaccination with live virus vaccines is not recommended for at least six weeks prior to the start of lymphodepleting chemotherapy, during Tecartus treatment, and until immune recovery following treatment with Tecartus.

Secondary Malignancies may develop. Monitor life-long for secondary malignancies. In the event that one occurs, contact Kite at 1-844-454-KITE (5483) to obtain instructions on patient samples to collect for testing.

Effects on Ability to Drive and Use Machines: Due to the potential for neurologic events, including altered mental status or seizures, patients are at risk for altered or decreased consciousness or coordination in the 8 weeks following Tecartus infusion. Advise patients to refrain from driving and engaging in hazardous activities, such as operating heavy or potentially dangerous machinery, during this period.

Adverse Reactions: The most common non-laboratory adverse reactions ( 20%) were fever, cytokine release syndrome, hypotension, encephalopathy, tachycardia, nausea, chills, headache, fatigue, febrile neutropenia, diarrhea, musculoskeletal pain, hypoxia, rash, edema, tremor, infection with pathogen unspecified, constipation, decreased appetite, and vomiting. The most common serious adverse reactions ( 2%) were cytokine release syndrome, febrile neutropenia, hypotension, encephalopathy, fever, infection with pathogen unspecified, hypoxia, tachycardia, bacterial infections, respiratory failure, seizure, diarrhea, dyspnea, fungal infections, viral infections, coagulopathy, delirium, fatigue, hemophagocytic lymphohistiocytosis, musculoskeletal pain, edema, and paraparesis.

About Kite

Kite, a Gilead Company, is a global biopharmaceutical company based in Santa Monica, California, with manufacturing operations in North America and Europe. Kites singular focus is cell therapy to treat and potentially cure cancer. As the cell therapy leader, Kite has more approved CAR T indications to help more patients than any other company. For more information on Kite, please visit http://www.kitepharma.com. Follow Kite on social media on Twitter (@KitePharma) and LinkedIn.

About Gilead Sciences

Gilead Sciences, Inc. is a biopharmaceutical company that has pursued and achieved breakthroughs in medicine for more than three decades, with the goal of creating a healthier world for all people. The company is committed to advancing innovative medicines to prevent and treat life-threatening diseases, including HIV, viral hepatitis and cancer. Gilead operates in more than 35 countries worldwide, with headquarters in Foster City, California.

Forward-Looking Statements

This press release includes forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995 that are subject to risks, uncertainties and other factors, including the ability of Gilead and Kite to initiate, progress or complete clinical trials within currently anticipated timelines or at all, and the possibility of unfavorable results from ongoing and additional clinical trials, including those involving Tecartus; the risk that physicians may not see the benefits of prescribing Tecartus for the treatment of blood cancers; and any assumptions underlying any of the foregoing. These and other risks, uncertainties and other factors are described in detail in Gileads Quarterly Report on Form 10-Q for the quarter ended March 31, 2022 as filed with the U.S. Securities and Exchange Commission. These risks, uncertainties and other factors could cause actual results to differ materially from those referred to in the forward-looking statements. All statements other than statements of historical fact are statements that could be deemed forward-looking statements. The reader is cautioned that any such forward-looking statements are not guarantees of future performance and involve risks and uncertainties and is cautioned not to place undue reliance on these forward-looking statements. All forward-looking statements are based on information currently available to Gilead and Kite, and Gilead and Kite assume no obligation and disclaim any intent to update any such forward-looking statements.

U.S. Prescribing Information for Tecartus including BOXED WARNING, is available at http://www.kitepharma.com and http://www.gilead.com .

Kite, the Kite logo, Tecartus and GILEAD are trademarks of Gilead Sciences, Inc. or its related companies .

View source version on businesswire.com: https://www.businesswire.com/news/home/20220722005258/en/

Jacquie Ross, Investorsinvestor_relations@gilead.com

Anna Padula, Mediaapadula@kitepharma.com

Source: Gilead Sciences, Inc.

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