Fetal Cell Lines Were Used to Make the Johnson & Johnson COVID VaccineHeres What That Means – Health.com

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Induced Pluripotent Stem Cells Reveal Causes of Disease – Technology Networks

Induced pluripotent stem cells (iPSC) are suitable for discovering the genes that underly complex and also rare genetic diseases. Scientists from the German Cancer Research Center (DKFZ) and the European Molecular Biology Laboratory (EMBL), together with international partners, have studied genotype-phenotype relationships in iPSCs using data from approximately one thousand donors.

Tens of thousands of tiny genetic variations (SNPs, single nucleotide polymorphisms) have been identified in the human genome that are associated with specific diseases. Many of these genetic variants are not located in the protein-coding regions of genes, but affect regulatory sections. Therefore, scientists are trying to find out if and in which tissues these variants can be linked to changes in the activity of specific genes.

Typically, such analyses are performed in blood cells or tissue biopsies, depending on the type of disease. "Pluripotent stem cells, however, might be better suited for this purpose in many cases, as they are undifferentiated and therefore reflect the ancestral state of all cells," says Oliver Stegle, division head at the German Cancer Research Center and group leader at EMBL. "Stem cells could be particularly relevant when searching for the cause of diseases that occur early in development." Pluripotent stem cells can be generated in the culture dish from normal body cells obtained from a blood sample, for example. They are referred to as "induced pluripotent stem cells," or iPSCs for short, since they are not naturally occurring stem cells.

Together with scientists from Stanford University and additional international cooperation partners, Oliver Stegle's team has compiled sequence and transcriptome data on iPSCs from around 1000 donors. The researchers systematically examined these data to identify correlations between individual genetic variants and altered expression patterns in stem cells. The results have now been published in the journal Nature Genetics.

For more than 67 percent of all genes active in iPSCs, the researchers found differential expression patterns depending on genetic variants. Many of these associations are novel and have not been described in somatic cell types before. For over 4000 of these associations, the genetic variants responsible for the altered expression patterns could be linked to specific diseases. These included, for example, variants associated with coronary heart disease, lipid metabolism disorders or hereditary cancers.

Stegle and colleagues also investigated whether iPS are suitable for identifying the causative genes of rare genetic diseases. They used iPSC lines from 65 patients who suffered from various rare diseases, whose causal gene defects were already known through previous analyses. In the transcriptome data of these iPSC lines, the scientists searched for particularly conspicuous "outliers" in the expression pattern. These analyses reliably led to the trace of the genetic basis of the disease. "Such screenings were previously impossible because there were simply no sufficiently large reference collections of iPS transcriptomes," explained Marc Jan Bonder, first author of the study.

"We were surprised to find such a large number of disease-associated genetic variants that are already visible in the expression pattern at the earliest time point of cell differentiation, represented by the iPSCs". Until now, the relevance of iPSCs for such biomedical analyses has been significantly underestimated.

In a companion paper, published in the same issue of Nature Genetics, Stegle and colleagues from EMBL-EBI and the Wellcome Trust Sanger Institute used more than 200 iPSC lines to investigate how genetic variants affect differentiation into neuronal cells.

The scientists performed RNA single-cell sequencing at different time points of neuronal cell differentiation. This allowed them to analyze how genetic variants affect expression patterns in different cellular states, including different neuronal cell types. "The study demonstrates the power of combining single-cell sequencing with iPSC technologies to dissect the effect of genetic variants in cell types that would otherwise be inaccessible," Stegle explains.

Reference:Bonder MJ, Smail C, Gloudemans MJ, et al. Identification of rare and common regulatory variants in pluripotent cells using population-scale transcriptomics. Nat Genet. 2021:1-9. doi:10.1038/s41588-021-00800-7

This article has been republished from the following materials. Note: material may have been edited for length and content. For further information, please contact the cited source.

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Pericardial Injection Effective, Less Invasive Way to Get Regenerative Therapies to Heart – NC State News

Injecting hydrogels containing stem cell or exosome therapeutics directly into the pericardial cavity could be a less invasive, less costly, and more effective means of treating cardiac injury, according to new research from North Carolina State University and the University of North Carolina at Chapel Hill.

Stem cell therapy holds promise as a way to treat cardiac injury, but delivering the therapy directly to the site of the injury and keeping it in place long enough to be effective are ongoing challenges. Even cardiac patches, which can be positioned directly over the site of the injury, have drawbacks in that they require invasive surgical methods for placement.

We wanted a less invasive way to get therapeutics to the injury site, says Ke Cheng, Randall B. Terry, Jr. Distinguished Professor in Regenerative Medicine at NCStates Department of Molecular Biomedical Sciences and professor in the NCState/UNC-Chapel Hill Joint Department of Biomedical Engineering. Using the pericardial cavity as a natural mold could allow us to create cardiac patches at the site of injury from hydrogels containing therapeutics.

In a proof-of-concept study, Cheng and colleagues from NCState and UNC-Chapel Hill looked at two different types of hydrogels one naturally derived and one synthetic and two different stem cell-derived therapeutics in mouse and rat models of heart attack. The therapeutics were delivered via intrapericardial (iPC) injection.

Via fluorescent imaging the researchers were able to see that the hydrogel spread out to form a cardiac patch in the pericardial cavity. They also confirmed that the stem cell or exosome therapeutics can be released into the myocardium, leading to reduced cell death and improved cardiac function compared to animals in the group who received only the hydrogel without therapeutics.

The team then turned to a pig model to test the procedures safety and feasibility. They delivered the iPC injections using a minimally invasive procedure that required only two small incisions, then monitored the pigs for adverse effects. They found no breathing complications, pericardial inflammation, or changes in blood chemistry up to three days post-procedure.

Our hope is that this method of drug delivery to the heart will result in less invasive, less costly procedures with higher therapeutic efficacy, Cheng says. Our early results are promising the method is safe and generates a higher retention rate of therapeutics than those currently in use. Next we will perform additional preclinical studies in large animals to further test the safety and efficacy of this therapy, before we can start a clinical trial.

I anticipate in a clinical setting in the future, iPC injection could be performed with pericardial access similar to the LARIAT procedure. In that regard, only one small incision under local anesthesia is needed on the patients chest wall, says Dr. Joe Rossi, associate professor in the division of cardiology at UNC-Chapel Hill and co-author of the paper.

The research appears in Nature Communications and was supported by the National Institutes of Health and the American Heart Association. Dr. Thomas Caranasos, director of adult cardiac surgery at UNC-Chapel Hill, also contributed to the work.

-peake-

Note to editors: An abstract follows.

Minimally invasive delivery of therapeutic agents by hydrogel injection into the pericardial cavity for cardiac repair

DOI: 10.1038/s41467-021-21682-7

Authors: Dashuai Zhu, Zhenhua Li, Ke Cheng, North Carolina State University; Thomas Caranasos, Joseph Rossi, University of North Carolina at Chapel HillPublished: March 3, 2021 in Nature Communications

Abstract:Cardiac patch is an effective way to deliver therapeutics to the heart. However, such procedures are normally invasive and difficult to perform. Here, we developed and tested a method to utilize the pericardial cavity as a natural mold for in situ cardiac patch formation after intrapericardial (iPC) injection of therapeutics in biocompatible hydrogels. In rodent models of myocardial infarction (MI), we demonstrated that iPC injection is an effective and safe method to deliver hydrogels containing induced pluripotent stem cells-derived cardiac progenitor cells (iPS-CPCs) or mesenchymal stem cells (MSCs)-derived exosomes. After injection, the hydrogels formed cardiac patch-like structure in the pericardial cavity, mitigating immune response and increasing the cardiac retention of the therapeutics. With robust cardiovascular regeneration and stimulation of epicardium-derived repair, the therapies mitigated cardiac remodeling and improved cardiac functions post MI. Furthermore, we demonstrated the feasibility of minimally-invasive iPC injection in a clinically-relevant porcine model as well as in human patients. Collectively, our study establishes iPC injection as a safe and effective method to deliver therapeutics to the heart for cardiac repair.

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Organoids of human lung and brain respond differently to SARS-CoV-2 – BioNews

1 March 2021

The body's response to infection by SARS-CoV-2 (the virus that causes COVID-19) varies across different cell types, according to a new study.

US researchers used organoids grown from human induced pluripotent stem cells to model how the virus affects cells of different organ systems. Although primarily a respiratory disease, COVID-19 can affect other organs in ways that are not fully understood, resulting in symptoms as diverse as arrhythmia, long-term fatigue and inability to think clearly.

'We're finding that SARS-CoV-2 doesn't infect the entire body in the same way. In different cell types, the virus triggers the expression of different genes, and we see different outcomes,' said research leader Professor Tariq Rana from the University of California San Diego School of Medicine.

Organoids are used as models to replicate some of the complexity of whole organs, although they lack immune cells and blood vessels. In this study they were used to model lung and brain tissues and, like their respective organs, they show ACE2 and TMPRSS2 receptors on the cell surface which are known bindingtargets for thevirus' 'spike proteins'.

Expression of ACE2 and TMPRSS2 on the surface of the lung and brain cells was explored by attaching a fluorescent marker to a non-infectious variant of the virus and exposing the tissues to it. The marker allowed the amount of binding of the pseudovirus spike proteins to be quantified which helped indicate the level of infectivity of thevirus in each tissue, showing lung organoids were much more susceptible.

The lung organoids had a ten-fold higher expression of ACE2 and TMPRSS2 proteins than the cells from the brain. Four different types of cells from the brain were tested, including cerebral organoids, and all showed lower levels of binding.

Differences in cell responses were also seen between the two types of tissue. The immune response in lung organoids involved expression of molecules known to mobilise immune cells: molecules such as interferons and cytokines. In contrast, brain organoids increased expression of molecules known to stimulate the innate immune response and support recognitions of pathogens. These molecules also support programmed cell death.

Professor Rana explained this could be significant as histeam previously saw a similar reaction in brain cells infected with Zika virus a virus known to stunt brain development in newborn babies.

The team also verified their findings by replicating the study using live SARS-CoV-2 virus in a biosafety laboratory specifically designed to handle high-risk infectious microbes.

The team are now supporting drug-discovery efforts by developing SARS-CoV-2 inhibitors and testing them on organoids derived from stem cells from donors of a variety of ethnic backgrounds.

The research was published in Stem Cell Reports.

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Study explores individual differences of SARS-CoV-2 infection in ACE2-expressing stem cells – News-Medical.Net

The clinical phenotype of coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is remarkable for its wide range of severity among individual patients. Genetic variations are known to mediate part of these differences.

To examine these differences, researchers in a new study used human induced pluripotent stem cells (iPSCs) from different genetically diverse individuals. These cells are used to model genetic disease since they contain the donors genetic information.

The study, published as a preprint on the bioRxiv* server, uses a panel of iPSCs from over 500 individuals. The researchers preferred undifferentiated iPSCs to reduce the time required to differentiate them, especially since infection is not always reliable.

Human iPSCs are not infected by SARS-CoV-2 since they express the viral entry receptor, the angiotensin-converting enzyme 2 (ACE2) at low levels. These cells do have high levels of the coronavirus receptor CD147 and of the serine protease TMPRSS2.

In order to render them susceptible to SARS-CoV-2 infection, the iPSCs were engineered with the relevant genes, inserted by adenovirus vectors (Ad), with almost 100% efficiency. These genes encoded human ACE2 and TMPRSS2 at over-expressing levels, leading to massive infection by the virus. TMPRSS2 overexpression alone was inadequate to produce this effect.

Two days post-infection of the ACE2-positive iPSCs, cell fusion was observed, with cell death after four days, in many cases. Under transmission electron microscopy (TEM), the infected cells showed characteristic changes, including zippered endoplasmic reticulum (ER), double-membrane spherules (DMS), and viral particles near the cell membrane.

Viral particles were seen within the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), along with double-membrane vesicles (DMVs. The latter is central to the synthesis of viral RNA and is absent from uninfected cells.

Thus, TEM is a useful tool to examine the lifecycle of this virus.

TEM images of infected ACE2-iPS cells (A) TEM images of infected ACE2-iPS cells. Zippered endoplasmic reticulum, double-membrane spherule (DMS) (B), and virus particles in the ERGIC (black arrows) (C) and near the cell membrane (black arrows) (D) were observed.

Gene and protein expression was assessed on day 3 post-infection in both infected and uninfected cells. This showed that infected cells had high levels of the viral genome and of ACE2, but not of undifferentiated cell markers or of innate immunity markers. The viral nucleocapsid (N) protein was also strongly expressed in the infected cells on day 2 post-infection.

RNA-seq analysis of these cells showed that in about 7% of all genes, there was a four-fold or more change in the level of expression, and none of these encoded markers of undifferentiation or innate immune markers.

Endoderm markers, except for the CER1 gene and for CD147, NRP1, and TMPRSS2 genes, known to be SARS-CoV-2-related markers, also remained unchanged. Mesoderm and ectoderm markers were also unchanged.

These results suggest that human iPS cells maintain an undifferentiated state even when SARS-CoV-2 replicates in large numbers.

In comparison to Vero cells, iPSCs proved to show stronger drug effects on SARS-CoV-2 infection, except for interferon-beta. Of the eight drugs tested, remdesivir showed maximum antiviral activity, and ivermectin showed high cytotoxicity.

Chloroquine and Favipiravir failed to suppress viral replication. The RNA-dependent RNA Polymerase (RdRp) inhibitors (Remdesivir and EIDD-2801) and TMPRSS2 inhibitors (Camostat and Nafamostat) were proved to have antiviral activity in these cells, indicating their value in drug evaluation

The researchers then infected iPSCs from eight different donors to explore differences in infection due to purely individual factors. The virus was replicated with varying efficiency among the eight donor cell lines, despite the comparable expression of ACE2 in all of them.

The virus showed greater replication capacity in male-origin cells than females, indicating the potential for sex-mediated differences in susceptibility to be replicated in these cells. Both the androgen receptor and its target TMPRSS2 gene, of which the latter is expressed at slightly higher levels in male iPSCs, may be implicated in this difference.

The viral lifecycle can be studied in these human iPSCs if the ACE2 receptor is overexpressed. The study showed the operation of sex-specific differences in the efficiency of infection and of viral replication. The inhibitory effects of drugs were also observable in this cell model.

These results suggest that by using our iPS cell panel, it will be possible to investigate the effects of race and blood type as well as gender on SARS-CoV-2 infection.

Furthermore, the identification of genetic mutations occurring at high frequency in cells that sustain infection is made possible by the use of iPSCs from a panel with documented genomic information.

While the infection efficiency varies among different donors, the fact that none of these donors had COVID-19 limits the inference that they had varying susceptibility to the disease. The study is being taken forward by using iPSCs from patients with mild and severe COVID-19, allowing findings in infected cells to be compared with the clinical presentation of the patient.

While mutations in certain innate immune genes may be associated with symptom severity, further research is required to understand how these genes function during COVID-19. This can be conveniently done in iPSCs, which allow the easy introduction of single nucleotide mutations.

It is noteworthy that the role of ACE2 overexpression has not been ruled out in the individual differences in SARS-CoV-2 infection within iPSCs from different donors. If so, another system employing ACE2-expressing somatic cells must be used to first assess the effect of the expression and mutation of ACE2 and other related genes.

Causes of individual differences, other than genetic, are not addressed by this system, including those due to advancing age and associated reductions in CD8 T cell cytotoxicity. Studies on blood specimens may help establish the cause of such variations in susceptibility and viral replication. The ACE2-positive iPSCs remain a useful platform to examine and decide on the cause of individual variations, identify high-risk groups and evaluate new drugs.

bioRxiv publishes preliminary scientific reports that are not peer-reviewed and, therefore, should not be regarded as conclusive, guide clinical practice/health-related behavior, or treated as established information.

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Study explores individual differences of SARS-CoV-2 infection in ACE2-expressing stem cells - News-Medical.Net

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Promoting Stem Cell Growth and Self-Renewal In Vitro – The Scientist

S

cientists grow and differentiate human embryonic and induced pluripotent stem (iPS) cells in laboratories around the world to study numerous cellular processes and develop regenerative medicines. To faithfully culture generations of these sensitive cells and drive them toward desired cell types, scientists add several components to their media and perform many steps, all while following strict protocols.

To continuously grow precious stem cells in vitro, researchers must preserve their stemness. They often accomplish this by using feeder cells, such as mouse or human fibroblasts, that deliver unknown factors that promote stem cell self-renewal. To grow in vitro, feeder cells require animal components such as fetal bovine serum (FBS). These ingredients do not comply with the Standard for Biological Ingredients, which regulates source materials derived from humans or animals. Additionally, feeder cells often carry viruses that infect stem cells. As an alternative, some scientists now use feeder-free cultures to avoid viruses and animal products in their cultures, and to reduce the labor involved with growing feeder cells. Cellular interactions within feeder-free media promote stem cell self-renewal.

Stem cells naturally interact with diverse extracellular matrix (ECM) proteins in their environment. The basement membrane alone is composed of numerous ECM proteins, which stem cells bind via integrin proteins. These interactions regulate stem cell fates. To replicate this cellular environment in the laboratory, scientists coat in vitro culture plates with ECM proteins and adhesion molecules to facilitate integrin binding. Some of the largest basement membrane proteins are laminins, which are heterotrimeric proteins composed of alpha, beta, and gamma chains.

Human stem cells express the 61 form of integrin, which binds to the laminin-511 isoform. The interaction of this integrin with human recombinant laminin-511 in cell culture promotes repeated stem cell renewal without feeder cells.1However, laminin-511 is an 800 kDa heterotrimeric protein, making it unsuitable for large-scale production and distribution.

To solve this problem, researchers produced a 150 kDa recombinant fragment of human laminin-511, ECMatrix-511 E8, that retained full integrin binding activity.2,3The scientists maintained several human embryonic and iPS cell lines in media containing the fragment. These cells retained high levels of pluripotency marker expression and could differentiate into all three germ layers. Because of its small size, ECMatrix-511 E8 is simple to produce at large-scale.

Compared to traditional basement membrane extracts, ECMatrix-511 E8 is more consistent lot-to-lot, promotes better cell adhesion and high proliferation rates, reduces cell clumping, and creates consistent microenvironments. The laminin fragment is ideal for CRISPR editing or clonal isolation applications because it allows for single cell passaging without the Rho-associated protein kinase (ROCK) inhibitor to prevent dissociation-induced apoptosis. Additionally, researchers can add the fragment directly to defined animal- and xeno-free media when passaging cells without pre-coating their cultureware. With ECMatrix-511 E8, scientists cultivate high quality feeder-free cultures to reliably propagate stem cells for regenerative medicine and beyond.

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Induced Pluripotent Stem Cells Market Growth, Competitive Landscape, Research Methodology, Business Opportunities, Statistics and Industry Analysis…

The latest report is the most recent study that offers 360 coverage of the Induced Pluripotent Stem Cells industry that has been facing the brunt of the adverse economic impact of the COVID-19 outbreak since the beginning of this year.

Top Companies Profiled in this Report are:

Thermo Fisher Scientific, Allele Biotechnology and Pharmaceuticals Inc., ABM (Applied Biological Materials Inc.), Addgene, Axol Bioscience, Cell Signaling Technology, Bluerock Therapeutics, Alstem LLC, Applied Stemcell Inc., ATCC, Creative Bioarray, Bristol-Myers Squibb, Bio-Techne, Reprocell Group Co., Primorigen Biosciences, ID Pharma Co. Ltd., Megakaryon Corp., FUJIFILM Cellular Dynamics, Inc., Waisman Biomanufacturing, Roslin Cell Sciences, Opsis Therapeutics, Corning Life Sciences, Fate Therapeutics, Genecopoeia, Gentarget Inc., Viacyte Inc., Ncardia, Invivogen, Lonza Group Ltd., Plasticell Ltd., Stemcell Technologies, Newcells Biotech, Orig3N Inc., Peprotech, Promega Corp., Promocell Gmbh, Qiagen N.V., System Biosciences Inc., Reprocell Inc., Sciencell Research Laboratories, MilliporeSigma, and Takara Bio Usa Inc.

Overview of the Induced Pluripotent Stem Cells report:

The Induced Pluripotent Stem Cells market has been broadly segmented to aid readers in gaining a more in-depth understanding of different facets and attributes of the market. The market scope for the new entrants and established companies has been assessed using various analytical tools, including the SWOT analysis, investment assessment, and Porters Five Forces analysis. Furthermore, into the study, the authors of the report have evaluated the financial standing of leading companies operating in the industry. They have provided crucial information on gross profits, revenue shares, sales volume, manufacturing costs, individual growth rate, and numerous other financial ratios of these competitors.

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Segmentation Analysis

The report gives an extensive analysis of various segments of the market by studying the product range, applications, major regions, and leading companies in the industry. Additionally, the report also dedicates an individual section to give a detailed analysis of the manufacturing process, which includes information collected through both primary and secondary sources of data collection. The primary source of data collection contains interviews of industry experts who offer accurate insights into the future market scenario.

Derived Cell Type Outlook (Revenue in USD Million; 20172027)

Application Outlook (Revenue in USD Million; 20172027)

End-User Outlook (Revenue in USD Million; 20172027)

Induced Pluripotent Stem Cellsmarket segmentation by geographical regions, the report has analysed the following regions-

North America (USA, Canada and Mexico)

Europe (Germany, France, UK, Russia and Italy)

Asia-Pacific (China, Japan, Korea, India and Southeast Asia)

South America (Brazil, Argentina, Columbia etc.)

Middle East and Africa (Saudi Arabia, UAE, Egypt, Nigeria and South Africa)

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The Induced Pluripotent Stem Cells Market Research/Analysis Report addresses the following questions:

WhichManufacturing Technologiesare prevalent in the production of Induced Pluripotent Stem Cells? What are the Recent Developmentsrelating to that technology? WhichTrendsare responsible for these developments?

Who are the leading vendors in the GlobalInduced Pluripotent Stem Cells Market? What are their individual market standing and contact information?

What is the current industrial scenario of the Global Induced Pluripotent Stem Cells Market? What were the Value, Volume, Production Capacity, Cost, and Profit Margin of the overall market?

What is the outcome of the competitive analysis on the Induced Pluripotent Stem Cells Market both in terms of companies and regions? What is the market assessment for the Induced Pluripotent Stem Cells Market as per the market segmented into types and applications?

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Induced Pluripotent Stem Cells Market Growth Opportunities, Analysis and Forecasts Report 2020-2026 with key players position (Fujifilm Holding…

The Induced Pluripotent Stem Cells Market grew in 2019, as compared to 2018, according to our report, Induced Pluripotent Stem Cells Market is likely to have subdued growth in 2020 due to weak demand on account of reduced industry spending post Covid-19 outbreak. Further, Induced Pluripotent Stem Cells Market will begin picking up momentum gradually from 2021 onwards and grow at a healthy CAGR between 2021-2025.

Deep analysis about Induced Pluripotent Stem Cells Market status (2016-2019), competition pattern, advantages and disadvantages of products, industry development trends (2019-2025), regional industrial layout characteristics and macroeconomic policies, industrial policy has also been included. From raw materials to downstream buyers of this industry have been analysed scientifically. This report will help you to establish comprehensive overview of the Induced Pluripotent Stem Cells Market

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The Induced Pluripotent Stem Cells Market is analysed based on product types, major applications and key players

Key product type:HepatocytesFibroblastsKeratinocytesAmniotic CellsOthers

Key applications:Academic ResearchDrug Development And DiscoveryToxicity ScreeningRegenerative Medicine

Key players or companies covered are:Fujifilm Holding CorporationAstellas PharmaFate TherapeuticsBristol-Myers Squibb CompanyViaCyteCelgene CorporationAastrom BiosciencesAcelity HoldingsStemCellsJapan Tissue EngineeringOrganogenesis

The report provides analysis & data at a regional level (North America, Europe, Asia Pacific, Middle East & Africa , Rest of the world) & Country level (13 key countries The U.S, Canada, Germany, France, UK, Italy, China, Japan, India, Middle East, Africa, South America)

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Key questions answered in the report:1. What is the current size of the Induced Pluripotent Stem Cells Market, at a global, regional & country level?2. How is the market segmented, who are the key end user segments?3. What are the key drivers, challenges & trends that is likely to impact businesses in the Induced Pluripotent Stem Cells Market?4. What is the likely market forecast & how will be Induced Pluripotent Stem Cells Market impacted?5. What is the competitive landscape, who are the key players?6. What are some of the recent M&A, PE / VC deals that have happened in the Induced Pluripotent Stem Cells Market?

The report also analysis the impact of COVID 19 based on a scenario-based modelling. This provides a clear view of how has COVID impacted the growth cycle & when is the likely recovery of the industry is expected to pre-covid levels.

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Can we freeze them? Yes we can, says Nkarta – Vantage

Paul Hastings, chief executive of Nkarta, is not shy about his companys achievement. We have mastered cryopreservation, he tells Evaluate Vantage.

And with that the company might have solved the biggest problem to have held back development of NK cell therapeutics: normally the cells cannot be frozen without risking changing their phenotype. Nkarta also reckons to have cracked another problem, transducing NK cells with a retrovirus, and its optimism matches the market mood of resurgent enthusiasm for these therapies.

That enthusiasm for NK cells took off at last years Ash meeting, and Nkarta, along with Gamida Cell, Fate Therapeutics and Nantkwest, celebrated surging stock prices. Remarkably, Nkarta presented no clinical data at the conference, though it has since started trials of its first Car-NK therapy, NKX101, targeting NKG2D ligands.

What do we want?

One of the things we defined right up front was what we wanted from a product, Mr Hastings says. Until we got that we werent going to try and move these things into the clinic.

For Nkarta a product must be off the shelf and cryopreserved though just how this is achieved is a proprietary secret and it must come in a vial, not a bag, so it can be multiple-dosed easily. The aim is to ship Car-NK cells just like an antibody directly to a doctor.

Thus affordability, a thorny issue for cell therapies, is at the core of Nkartas thinking. Mr Hastings reckons cost of manufacturing can be limited to $2,000 per vial of a billion NK cells; and, as each dosing cycle comprises three lots of 100 million cells, each vial could give up to three cycles.

As for supply, Nkarta sources NK cells from healthy donors, in contrast to Fate, which uses induced pluripotent stem cells (iPSCs). Mr Hastings says each healthy donor can yield 500 doses, and manufacturing, including standard viral transduction, takes 14 days, with the inclusion of membrane-bound IL-15 improving persistence and reducing exhaustion, ensuring that cells are maximally potent when delivered.

TheiPSC approach is seen as more advanced, but iscomplex asdesired characteristics need to be engineered in. Mr Hastings says with donor-derived cells you start with a large number of cells, and theyre actually NK cells, though he does not rule out looking at iPSCs in future.

Were all placing bets, and this is our bet, adds James Trager, Nkartas chief scientific officer. And he denies that donor-derived cells lack flexibility, saying the donor pool is very diverse: There are a fair number of donors walking around who have NK cells with pretty exceptional properties.

For now, however, Nkarta is not looking at donors with NK cells expressing high-affinity CD16; Fate, for instance, engineers this into most of its projects to improve antibody-dependent killing.

Clinical data

Investors do not have long to wait for the first evidence of NKX101s clinical activity. Mr Hastings says Nkarta expects interim results from a handful of patients at a medical meeting not by press release towards the end of 2021.

And an IND filing for the groups second asset, a CD19-targeting Car-NK coded NKX019, is due in the current quarter. This gives Nkartas pipeline a mix of proven (CD19) and high-risk (NKG2D ligands) approaches, and final proof-of-concept data for both are due in the first half of 2023; current cash, meanwhile, takes Nkarta to the second half of that year.

On a design level both therapies differ from current Car-T approaches in using as the co-stimulatory domain not 4-1BB or CD28 but Ox40, which Mr Trager says gives NK cells the best serial killing activity.

Even though the CD19 space is extremely crowded, unlike many other cell therapy players Nkarta is not using a CD19 approach just to test its tech. We absolutely believe that a CD19 programme thats allogeneic, cryopreserved in a vial is going to be very competitive, stresses Mr Hastings.

And the groups next project will target solid tumours, but for now remains under wraps. The way well disclose targets is when we have a clinical candidate, says the chief exec.

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Can we freeze them? Yes we can, says Nkarta - Vantage

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Global Induced Pluripotent Market Positive Outlook, Revenue Generation & Leading Manufacturers, Forecast 2026||CELGENE CORPORATION; Astellas…

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Few of the major competitors currently working in the induced pluripotent market areBristol-Myers Squibb Company; CELGENE CORPORATION; Astellas Pharma Inc.; Thermo Fisher Scientific; Cell Applications, Inc.; Axol Bioscience Ltd.; Organogenesis Holdings; Merck KGaA; FUJIFILM Holdings Corporation; Fate Therapeutics; KCI Licensing, Inc.; Japan Tissue Engineering Co., Ltd.; Vericel; ViaCyte, Inc.; STEMCELL Technologies Inc.; Horizon Discovery Group plc; Lonza; Takara Bio Inc.; Promega Corporation and QIAGEN.

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Global Induced Pluripotent Market Positive Outlook, Revenue Generation & Leading Manufacturers, Forecast 2026||CELGENE CORPORATION; Astellas...

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