Category Archives: Stem Cells

Global Synthetic Stem Cells Market is valued at approximately USD $billion in 2021 and is anticipated to grow with a healthy growth rate of more than % over the forecast period 2022-2028.

Stem cell therapies work by promoting endogenous repair that is, they help damaged tissue in repairing itself by secreting paracrine factors, including proteins and genetic materials. While stem cell therapies can be effective, they are also associated with some risks of both tumor growth and immune rejection. The increasing incidences of various cardiovascular diseases and government investment in research & development activities have led to the adoption of Synthetic Stem Cells across the forecast period.

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For Instance: as per the WHO, An estimated 17.9 million people died from CVDs in 2019, representing 32% of all global deaths. Of these deaths, 85% were due to heart attack and stroke. Also, with the development of stem cells for stem line banking, the adoption & demand for Synthetic Stem Cells is likely to increase the market growth during the forecast period. However, unclear and unstructured regulations impede the growth of the market over the forecast period of 2022-2028.

The key regions considered for the Global Synthetic Stem Cells Market study include Asia Pacific, North America, Europe, Latin America and Rest of the World. North America is the leading region across the world in terms of market share owing to the growing investment in research and development activities. Whereas, Asia-Pacific is also anticipated to exhibit the highest growth rate over the forecast period 2022-2028. Factors such as rising population, rising incidences of injuries and improving healthcare infrastructure would create lucrative growth prospects for the Synthetic Stem Cells Market across Asia-Pacific region.

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Major market players included in this report are:

The objective of the study is to define market sizes of different segments & countries in recent years and to forecast the values to the coming eight years. The report is designed to incorporate both qualitative and quantitative aspects of the industry within each of the regions and countries involved in the study. Furthermore, the report also caters the detailed information about the crucial aspects such as driving factors & challenges which will define the future growth of the market. Additionally, the report shall also incorporate available opportunities in micro markets for stakeholders to invest along with the detailed analysis of competitive landscape and product offerings of key players. The detailed segments and sub-segment of the market are explained below:

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By Application:

Cardiovascular Disease

Neurological Disorders

Cancer

Diabetes

Gastrointestinal

Musculoskeletal Disorder

By End Use:

Hospitals and Surgical Centers

Academic Institutes

Research Laboratories

Pharmaceutical and Biotechnology Companies

Others

By Region:

North America

U.S.

Canada

Europe

UK

Germany

France

Spain

Italy

ROE

Asia Pacific

China

India

Japan

Australia

South Korea

RoAPAC

Latin America

Brazil

Mexico

Rest of the World

Table of Content

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Synthetic Stem Cells Market 2022 Size, Share, Future Plans, Competitive Landscape and Forecast to 2030 This Is Ardee - This Is Ardee

DUBLIN, July 25, 2022 /PRNewswire/ -- The 'Global Research Antibodies & Reagents Market by Product (Antibodies (Type, Form, Source, Research Area), Reagents), Technology (Western Blot, Flow Cytometry, ELISA), Application (Proteomics, Genomics), End-user (Pharma, Biotech, CROs), and Region - Forecast to 2027'report has been added to ResearchAndMarkets.com's offering.

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The global research antibodies and reagents market is projected to reach USD 16.1 billion by 2027 from USD 11.6 billion in 2022, at a CAGR of 6.7%

The research antibodies and reagents market evolved owing to factors such as increasing proteomics and genomics research, growing demand for antibodies for research reproducibility, and increasing R&D expenditure in the life sciences industry.

Driven by the increasing demand for personalized medicine and structure-based drug design. It is expected that the global research antibodies and reagents market will witness significant growth in the coming years.

On the basis of product, the reagents segment holds the highest market share during the forecast period

On the basis of product, the research antibodies and reagents market are segmented into reagent and antibodies. In 2021 the reagent segment accounted for the larger market share. Factors such as increasing applications of biosciences and biotechnology within the healthcare and pharmaceutical fields is driving the market.

On the basis of technology, the flow cytometry segment is expected to register the highest CAGR during the forecast period

On the basis of technology, the research antibodies and reagents market is segmented into western blotting, flow cytometry, ELISA, Immunohistochemistry, Immunofluorescence, Immunoprecipitation, and other technologies. During the forecast period the flow cytometry segment is expected to witness the highest growth.

Factors such as advantages of this technique, its ability to perform simultaneous multi-parameter analysis on single cells within a heterogeneous mixture, offering high throughput along with technological innovations in flow cytometry and increasing oncology research, are driving the growth of this segment.

On the basis of application, the proteomics segment holds the highest market share during the forecast period

On the basis of application, the research antibodies and reagents market is segmented into proteomics, drug development and Genomics. In 2021, Proteomics held the largest share of the global research antibodies and reagents market. Factors such as increasing efficiency maps drug-protein and protein-protein interactions.

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Additionally, proteomic technologies have minimized the cost, time, and resource requirements for chemical synthesis and biological testing of drugs and are highly efficient. Such factors are driving the market.

On the basis of end-user, the pharmaceutical & biotechnology segment holds the highest market share during the forecast period

The research antibodies and reagents market is divided into the pharmaceutical & biotechnology companies, academic & research institutions and Contract Research Organizations.

In 2021 the pharmaceutical & biotechnology companies held the largest share of the global research antibodies and reagents end-user market. Factors such as growing use of research antibodies in drug development for the identification and quantification of biomarkers and other techniques are driving the market.

By Region, Asia Pacific is expected to register the highest CAGR during the forecast period

During the forecast period (2022 to 2027), the Asia Pacific research antibodies and reagents market is expected to grow at the highest CAGR. Factors such as increasing research in proteomics and genomics and growing research funding, investments by pharmaceutical and biotechnology companies, and growing awareness in the region are driving the market in the region.Key Players

The key players operating in the research antibodies and reagents systems include Thermo Fisher Scientific, Inc. US), Merck KGaA (Germany), Abcam plc (UK), Becton, Dickinson and Company (US), Bio-Rad Laboratories (US), Cell Signaling Technology (US), F. Hoffmann-La Roche (Switzerland), Danaher Corporation (US), Agilent Technologies (US), PerkinElmer (US), Lonza (Switzerland), GenScript (China), and BioLegend (US).

Premium Insights

Increasing R&D Expenditure in the Life Science Industry to Drive Market Growth

Proteomics Accounted for the Largest Share of the Asia-Pacific Research Antibodies and Reagents Market in 2021

China Shows the Highest Revenue Growth Opportunities During the Forecast Period

North America Will Continue to Dominate the Research Antibodies and Reagents Market Until 2027

Developing Markets to Register a Higher Growth Rate in the Forecast Period

Market Dynamics

Drivers

Restraints

Opportunities

Emerging Markets

Personalized Medicine and Protein Therapeutics

Growth in Stem Cell and Neurobiology Research

Increasing Focus on Biomarker Discovery

Rising Interest in Outsourcing

Challenges

Industry Trends

Increasing Research on Therapeutic Antibodies

Recombinant Antibodies Supporting the Smooth Transition from in Vitro to in Vivo

Growing Consolidation of the Life Sciences Market for Antibodies and Reagents

Stakeholder Analysis

Impact of COVID-19 on the Research Antibodies and Reagents Market

Supply Chain Analysis

Technology Analysis

Regulatory Analysis

Porter's Five Forces

Companies Mentioned

Abcam plc

Agilent Technologies, Inc.

Analytik Jena AG

Atlas Antibodies

BD

Bio-Rad Laboratories, Inc.

Biolegend

Cell Signaling Technology, Inc.

Danaher Corporation

Dovetail Genomics

F. Hoffmann-La Roche

Fujirebio Diagnostics AB

Genscript

Illumina, Inc.

Immunoprecise Antibodies Ltd

Lonza

Merck KGaA

Omega Bio-Tek

Perkinelmer, Inc.

Thermo Fisher Scientific, Inc.

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Global Research Antibodies & Reagents Markets, 2022-2027 - Growth in Stem Cell and Neurobiology Research / Increasing Focus on Biomarker Discovery...

Scientists from the RIKEN Center for Integrative Medical Sciences in Japan in collaboration with other researchers from around the world have discovered that recombinations of specific genomic sequences that are repeated millions of times in the genome of each of our cells are pervasively found in both normal and in disease states. Identifying the mechanisms that lead to this myriad of recombinations involving DNA sequences that were once considered as junk, may be crucial to understanding how our cells develop and what can make them unhealthy.

Following the discovery of DNA, it was long believed that all the cells in our body share the same genetic code, safely guarded within the nucleus. However, modern advances in DNA sequencing have challenged this view: we now know that mutations accumulate in the genome of single cells starting from the very early stages of development. However, the magnitude of this phenomenon and how it contributes to disease is not well understood.

In this work, published in Cell, the authors looked at certain repeated genomic sequences, called Alu and L1, and developed a method to study these specific sequences of DNA that are repeated millions of times in the genome of each cell. It was already known that they recombine with each other, generating mutations often found in cancer and other genetic disorders. By analyzing the DNA of donors unaffected by disease, the researchers identified millions of DNA mutations caused by the recombination of these repeated sequences, and further discovered that different tissues in the body are characterized by different recombination signatures.

The researchers also found that the differentiation of human stem cells into neuronal cells is accompanied by distinct changes of recombination of repeat sequences. This indicates that this particular type of DNA mutation may be a physiological phenomenon involved in human development.

Finally, the researchers looked at the recombination of repeated sequences in samples from people affected by Alzheimer's and Parkinson's disorders, the two most prominent neurodegenerative disorders in the developed world. They found signatures of recombination that are specific to each disease, suggesting that genomic recombinations caused by these repeated sequences are involved in brain diseases.

According to Giovanni Pascarella, first author of the study, "We have shown in this study that the recombination of repeat elements in the human genome is a widespread phenomenon that contributes to the complex constellation of genomic variants making up our genomes."

According to, Piero Carninci, Principal Investigator and co-corresponding author of the study, "We hypothesize that it might be that random recombinations of Alu and LI in somatic cells may occasionally prime the genome of individual cells at vulnerable sites and drive the transition from healthy to pathological states."

"However," he continues, "what is difficult to know at this point is to determine whether the recombinations in disease are truly causative or if they are effects of the disease state. Further studies need to be done to understand this important question."

Experimental study

Cells

Recombination of repeat elements generates somatic complexity in human genomes

25-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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DNA recombinations are widespread in human genomes and are implicated in both development and disease - EurekAlert

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,

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

Cerebrospinal fluid (CSF) from young mice improved the ability of older mice to recall recent memories, according to an NIA-funded study published in Nature. The findings suggest that at least one CSF protein may trigger the growth and maturation of cells that help protect brain function in older mice. The study results appear to underscore the therapeutic potential of CSF proteins found in young mice fluid as well as the role that these helper cells play in brain aging.

CSF is a clear fluid that surrounds the brain and spinal cord, providing protection as well as vital nutrients, including proteins, fats, sugars, and vitamins. The composition of CSF changes with age, and it is not clear whether these changes contribute to age-related memory loss.

Previously, scientists discovered that blood from young mice may rejuvenate the brains of older mice. In this more recent study, an international team of scientists from Stanford University; Palo Alto Veterans Institute for Research; Saarland University, Germany; University of Gothenburg, Sweden; and University College London Queen Square Institute of Neurology explored whether CSF from young mice might have similar effects.

To address this possibility, the researchers first tested whether CSF from young mice influenced the ability of older mice to recall memories of foot shocks. Mice are considered adults at two months old and have a life expectancy of approximately two years. First, a group of 20-month-old mice was trained with foot shocks to freeze in motion when presented with a light and sound cue. Then CSF, drawn from three-month-old mice, was injected into the brains of some of the older mice. The mice that received the CSF froze in motion more often when presented with the cue than those that received a control fluid, suggesting the young CSF helped the older mice remember better.

Next, the researchers searched for clues to how this happened by examining the brains of the older mice. Their results suggest that the young CSF caused stem cells, called oligodendrocyte progenitor cells (OPCs), to multiply and mature. Mature oligodendrocytes help neurons by creating myelin, which is a waxy material that insulates long, stringy axons, a part of neurons that relays signals to other cells. Much of this was observed in the hippocampus, a region of the brain involved in memory. Other studies of mice have also found links between brain myelin and memory.

Further experiments revealed that a protein called Fgf17 may have played a critical role in improving memory and triggering OPC growth. Mice injected with Fgf17 performed better on the foot shock memory tests and had greater OPC growth and maturation than those that received a control solution. In contrast, mice treated with a blocker of Fgf17 activity performed worse on the memory tests than those receiving a control drug. Moreover, the Fgf17 blocker inhibited the OPC growth in the presence of young CSF.

The results support the idea that the CSF and myelinating helper cells may play a critical role in the aging brain. Furthermore, by studying Fgf17 and other factors found in younger CSF, scientists may discover new clues to treating age-related brain disorders.

This research was supported in part by NIA grants RF1-AG064897-02 and T32AG000266.

Reference: Iram T, et al. Young CSF restores oligodendrogenesis and memory in aged mice via Fgf17. Nature. 2022;605(7910):509-515. doi: 10.1038/s41586-022-04722-0.

Read more here:
Cerebrospinal fluid from young mice improved memory in older mice - National Institute on Aging

Bhubaneswar, July 26: A 30-year-old woman suffering from a very rare blood disorder has been successfully treated at the Institute of Medical Sciences and SUM Hospital recently.

The woman, hailing from Jajpur, was admitted into the hospital in the last week of May for anemia and bleeding requiring repeated blood and platelet transfusions spread over one month.

The patients bone marrow, the site of blood cell formation, was biopsied for diagnosing the disease and investigations revealed that she was suffering from Severe Aplastic Anemia, a rare blood disorder, where the bone marrow was found completely empty and unable to form new blood cells, Dr. Priyanka Samal, Head of the department of Hematology in the hospital, said.

Such patients have very low hemoglobin, low total white blood cell count and very low platelet count, she said adding they complain of severe weakness, bleeding from orifices or menorrhagia which does not stop unless platelets are transfused.

These patients also get infected frequently requiring hospitalization and administration of intra-venous antibiotics and often succumb to sepsis, Dr. Samal said.

She said the very severe form of Aplastic Anemia had a very poor prognosis and such patients survived only a few months. The incidence of this disease is higher in Asia compared to the west and the only cure is available through stem cell transplantation, she added.

The woman underwent stem cell transplantation on July 5 with stem cells donated by her 28-year-old brother though they had a major blood group discrepancy which was taken care of very delicately, Dr. Samal said while pointing out that stem cells in the donors body get replaced within 4-6 weeks without any adverse effect to the donor.

A growth factor of 4-5 days is required for the stem cells to get mobilized from the bone marrow to peripheral blood. Following this, apheresis was done for 4-5 hours for harvesting only stem cells from the blood and it was just like a normal platelet donation process, she said.

The patient was then subjected to high dose chemotherapy after which the donors stem cells were infused into her body like normal blood transfusion. It took 12-13 days for these donor cells to make the new blood cells in the patients body.

The woman, whom the disease might not have given a few months to live, was discharged from the hospital on July 23 in good condition and full blood count recovery though she needs to be closely observed for the next few months for complete immunological recovery, Dr. Samal said.

The patient can now hope to lead an uneventful life, at least from this ailments aspect, she said.

She also thanked the Medical Superintendent Prof. Pusparaj Samantasinhar, Additional Medical Superintendent Dr. Rajesh Lenka, the department of transfusion medicine, laboratory and radiology teams as also the nursing staff whose tireless efforts led to the success of the procedure.

Continue reading here:
Patient with rare blood disorder successfully treated at SUM - Odisha News In English

Back in the early 1990s, when 30,000 eager attendees filed into the Society for Neurosciences annual meeting, the talk of the conference was Samuel Weisss jaw-dropping discovery of neural stem cells. At the time, scientific dogma insisted the brain had no regenerative capacity: you do drugs, you bump your head, you lose neurons and they never come back. Weisss research showed that cells in the adult brain could make new neurons, opening the door to dramatic possibilities for neural repair.

There was just one hitch.

They had kind of taken this melon-ball scoop of the brain, so they didnt really know where the stem cells were, says Cindi Morshead, who arrived at the conference as a University of Toronto graduate student and full-fledged neuro nerd.

Her research, however, focused on the lateral ventricles of the forebrain, and she had a sneaking suspicion thats where they were hiding. I just walked up to Sams graduate student and said, I know where the stem cells are. A handful of experiments later, she was proved right.

Morshead wasnt a brash student, brimming with confidence. In fact, as one of just five female scientists currently at U of Ts Donnelly Centre for Cellular and Biomolecular Research, she says she still contends with impostor syndrome. But ever since taking a third-year undergraduate course in neuro-psychopharmacology where she first became enamoured with the brain Morshead has seized any opportunity to work in the field she loves.

I kicked ass on this one neuroanatomy exam, and that meant I got to do research with Derek van der Kooy, who ended up being my mentor, she recalls. Her graduate work involved using viral vectors for cell lineage tracing, a new technique at the time.

There were people who needed the expertise that I had people like Weiss and Bryan Kolb, both titans in Canadian neuroscience so I literally called them up and said, I know this is what you should do, and I can do it for you, Morshead says. The whole idea of putting myself out there was hard, but I really had no choice. I had to break into that field.

Now the chair of the anatomy division in the department of surgery at U of T, Morshead is once again helping upend expectations about brain cells. As part of the regenerative medicine hub Medicine by Design, which receives funding from the Canada First Research Excellence Fund, shes exploring the possibility of using gene therapy to treat neurodegenerative disorders, particularly strokes.

For the 50,000-plus Canadians who have a stroke each year, cutting off the blood supply to their brains, time is of the essence. Neurons are very greedy cells and require constant oxygen and glucose, so they start to die within minutes to hours, she says. A stroke can often cause certain brain cells called astrocytes which play a key role in supporting the transmission of neural messages and keeping the brain in equilibrium to become toxic, killing even more neurons.

What if, Morshead and her team wondered, we could make new neurons in the brain to take their place? To do so, they infected astrocytes in mice brains with a nonreplicating virus containing the DNA code for NEUROD1, a transcription factor that expresses genes typically found in neurons. Within a matter of weeks, those astrocytes had transformed into functioning neurons. Were able to replace a lost cell and were able to get rid of a toxic cell, Morshead says. Its sort of a double whammy.

Following a stroke, people tend to have motor impairments and weakened grip strength. But Morshead found that after the mice were given the transcription factors, all of these behaviours improved, and it was correlated with this new production of neurons, she says. So that was phenomenal.

It points to the possibility of treatment that might profoundly help stroke patients, who are often given very little reason to hope. Even small changes can have an enormous impact on the quality of their lives. Being able to pick up a utensil or scratch the itch on their neck rather than asking someone else to scratch it that can make such a difference, she says.

This gene therapy has implications for other neurodegenerative diseases, too: Morshead is collaborating with Sunnybrook scientists like Carol Schuurmans (whose group studies ALS) and JoAnne McLaurin (whose team works in Alzheimers disease). Theyre finding that using gene therapy to turn astrocytes into neurons is showing improved outcomes in their animal models, which is huge.

When Morshead first pursued brain research as an undergraduate student, she told herself shed just keep doing the work until she didnt like it anymore and then maybe shed go to teachers college or try dentistry. But Ive been really lucky, she says. My work has taken many turns, but I still love the brain.

Four ways tech can help diagnose and manage neurodegenerative diseases

Disclaimer This content was produced as part of a partnership and therefore it may not meet the standards of impartial or independent journalism.

See the rest here:
Meet the U of T neuroscientist helping the brain heal itself - Toronto Star

Researcher Dhruv Sareens own stem cells are now orbiting the Earth. The mission? To test whether theyll grow better in zero gravity.

Scientists at Cedars-Sinai Medical Center in Los Angeles are trying to find new ways to produce huge batches of a type of stem cell that can generate nearly any other type of cell in the body and potentially be used to make treatments for many diseases. The cells arrived over the weekend at the International Space Station on a supply ship.

I dont think I would be able to pay whatever it costs now" to take a private ride to space, Sareen said. At least a part of me in cells can go up!"

The experiment is the latest research project that involves shooting stem cells into space. Some, like this one, aim to overcome the terrestrial difficulty of mass producing the cells. Others explore how space travel impacts the cells in the body. And some help better understand diseases such as cancer.

By pushing the boundaries like this, its knowledge and its science and its learning, said Clive Svendsen, executive director of Cedars-Sinais Regenerative Medicine Institute.

Six earlier projects from the U.S., China and Italy sent up various types of stem cells including his teams study of the effects of microgravity on cell-level heart function, said Dr. Joseph Wu of Stanford University, who directs the Stanford Cardiovascular Institute. Wu helped coordinate a series of programs on space-based stem cell research last year.

Earthly applications of much of this research may be a little ways off.

At this point, the only stem cell-based products approved by the Food and Drug Administration contain blood-forming stem cells from umbilical cord blood for patients with blood disorders such as certain cases of lymphoma. There are no approved therapies using the kind of stem cells being sent to space or others derived from them, said Jeffrey Millman, a biomedical engineering expert at Washington University in St. Louis.

But clinical trials underway involving stem cells target conditions such as macular degeneration, Parkinsons disease and heart attack damage. And Millman is involved in research that could lead to a new approach for treating Type 1 diabetes.

Scientists see great promise in stem cells.

THE GRAVITY DILEMMA

That promise is tempered by a frustrating earthly problem: The planets gravity makes it tough to grow the vast quantities of cells necessary for future therapies that may require more than a billion per patient.

With current technology right now, even if the FDA instantly approved any of these therapies, we dont have the capacity to manufacture what's needed, Millman said.

The issue? In large bioreactors, the cells need to be stirred vigorously or they clump or fall to the bottom of the tank, Millman said. The stress can cause most cells to die.

In zero G, theres no force on the cells, so they can just grow in a different way, Svendsen said.

The Cedars-Sinai team has sent up what are called induced pluripotent stem cells. Many scientists consider them the perfect starting materials for all sorts of personalized, cell-based treatments. They carry a patients own DNA, and their versatility makes them similar to embryonic stem cells, only they are reprogrammed from adults' skin or blood cells.

For their experiment, which is being funded by NASA, a shoebox-sized container holds bags filled with spheres of cells and all of the pumps and solutions needed to keep them alive for four weeks. The cargo will also include neural stem cells originating from Svendsen. The scientists used stem cells derived from their own white blood cells because it was easy for them to give consent.

They will run the experiment remotely with a box of cells on Earth for comparison. They'll get the space experiment back in five weeks or so, when it returns in the same SpaceX capsule.

The work is designed to pave the way for more NASA-funded research. If they are able to figure out how to make billions of cells in orbit, Svendsen said, the impact could be huge.

A HIGH-FLYING FUTURE

During the same cargo launch, researchers from the University of California, San Diego, sent blood stem cells to the space station, a repeat of an experiment they did last year. They want to find out if low Earth orbit induces faster aging in the cells, leading to problems that set the stage for precancerous changes. One goal is to protect astronauts' health.

Afshin Beheshti, a researcher at NASA Ames Research Center, said scientists are just beginning to understand some of the risks of space travel.

Theres more unknowns in space than there are knowns," he said. Any new type of experiment is going to shed light on how the body responds to the space environment.

Ultimately, Beheshti said, the research should yield more than practical, earthly solutions like new medicines. It will also help with far-off human aspirations, like living on other planets.

The Associated Press Health and Science Department receives support from the Howard Hughes Medical Institutes Department of Science Education. The AP is solely responsible for all content.

Excerpt from:
High-flying experiment: Do stem cells grow better in space? - ABC News

For the past seven weeks, four-year-old Minh Nguyen has spent most of her time in hospital, receiving regular blood transfusions because her doctors have not been able to find acompatible stem cell donor.

Minh was diagnosed with bone marrow aplasia, arare blood disease that affects her bone marrow's ability to generate white blood cells. She hasspent the last three weeks in hospital full-time.

"It's been hard for her because she doesn't have the life of a four-year-oldchild anymore," said Minh's mother, Diem Nguyen.

Nguyen was speaking at an event organized for Minh at a Montreal restaurant Saturday, where dozens of people with similar genetic makeups to Minh showed up to volunteer as potential stem cell donors.

There is an international registry of 25millionpeople who have provided consent and genetic information in order to donate stem cells, which come from the bone marrow of a healthy adult.

But about 70 per cent of those on the registry are white, putting donor recipients of from other ethnic backgroundsat a significant disadvantage.

Before Saturday's event, Minh's mother described her daughter's chances as one in a million.

Marie-Cindel Surprenant says she felt a personal connection to Minh Nguyen's story when learned about it from a colleague.

"I think that if somehow I can help her, I'll do it, so I think it's a good purpose in my life," she said.

Surprenant is half-Asian, half-white, like Minh.

The toddler's condition can be treated with stem cells from a compatible donor but because of the lack of diversity in the global donor registry, finding a match has been incredibly difficult.

Nguyen said she was moved by the amount of people who came to get swabbed and show their support on Saturday.

"I'm really grateful. These are complete strangers coming to help Minh," she said.

Nguyen said her daughter has shown incredible resilience throughout her time in hospital.

"She's laughing, she's reading, she's playing as much as she can," the mother said.

Friday evening, Nguyen said she started crying at the thought "that she might not outlive me, in terms of lifespan" and Minh asked her why she was sad.

"I said, 'No, I'm just having a bit of allergies,' and she said, 'Please, don't say that, mom, I know you're crying.' I said, 'I just want to stay with you as long as I can.' And she said, 'I'm here for you mom, I'm not leaving you,'" Nguyen recounted.

"She is incredible."

Samuel Sassine, a University of Montreal medical school student who attended Saturday's event, works for a foundation called Swab the World, which was founded by Mai Duong, a woman of Vietnamese descent who also struggled to find a stem cell donor, to encourage people from diverse ethnic backgrounds to become donors.

Sassine said there is currently no one in the world registered with the same DNA background as Minh.

"White people have more chances of living through this disease than other people from other ethnic backgrounds and thatis unacceptable," he said.

"In 2022, ethnic non-equity should not exist, and to live or to not live based on your skin colour should not exist either."

Alex Fong, wholike Surprenant, came to get swabbed on Saturday to see if he could be a potential donor, said Minh's story had move him.

"When I found out that not a lot of Asian donors were doing stem cells, I thought it would be a good idea to come and just participate," said Fong.

Sassinesays he hopes to see more awareness spread in schools about theof the lack of diversity in stem cell donor registries.

He encourageseveryone who is eligible, regardless of ethnicity, to register as a stem cell donor.

In Quebec, stem cell donor registrations are carried out through Hma-Qubec.It involves filling in a questionnaire online and then swabbing cells in your mouth with a kit sent by mail. Once you send it back, you can be part of the donor bank, Sassine explained.

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Stem cell donors volunteer in hopes of saving Montreal toddler with rare blood disease - CBC.ca

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 -...