2023 Stem Cell Therapy Market Key Trends: Dynamics Shaping the … – Digital Journal

PRESS RELEASE

Published April 11, 2023

Stem Cell Therapy Market Report 2023 | 125 Pages Latest Research | provides all-inclusive update on growth segments, recent developments of top players [Ernst & Young, Aon Consulting, McKinsey & Company, Accenture, PriceWaterhouseCoopers (PwC), Affiliated Computer Services, KPMG Consulting] with their revenue and CAGR status. It also covers various marketing strategies of top manufacturers.

"Final Report will add the analysis of the impact of COVID-19 on this industry."

Global Stem Cell Therapy Market Report 2023-2028 is a comprehensive analysis of the latest trends and opportunities in the industry. The report presents a detailed assessment of the market size, growth factors, and demand status, as well as the challenges and risks faced by businesses across all geographic regions. It also features the latest technological advancements, SWOT, and PESTLE analysis, providing a holistic overview of the industry's competitive landscape and regional segments. This insightful report covers key players profiling with their company profiles, supply-demand scope, and global trending technologies.

Additionally, Stem Cell Therapy market (125 Pages) report is a valuable addition to any company's future strategies and path forward, as it offers a deep understanding of the industry revenue and competitive landscape. With its precise information and thoughtful analysis, the report provides businesses with the necessary tools to make informed decisions and stay ahead of the competition.

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The global Stem Cell Therapy market size was valued at USD 12932.56 million in 2022 and is expected to expand at a CAGR of 8.2% during the forecast period, reaching USD 20753.25 million by 2028.Stem-cell therapy is the use of stem cells to treat or prevent a disease or condition.Bone marrow transplant is the most widely used stem-cell therapy, but some therapies derived from umbilical cord blood are also in use. Research is underway to develop various sources for stem cells, and to apply stem-cell treatments for neurodegenerative diseases and conditions, diabetes, heart disease, and other conditions.With the ability of scientists to isolate and culture embryonic stem cells, and with scientists' growing ability to create stem cells using somatic cell nuclear transfer and techniques to create induced pluripotent stem cells, controversy has crept in, both related to abortion politics and to human cloning. Additionally, efforts to market treatments based on transplant of stored umbilical cord blood have been controversial.

Key Players covered in the global Stem Cell Therapy Market are:

The report focuses on the Stem Cell Therapy market size, segment size (mainly covering product type, application, and geography), competitor landscape, recent status, and development trends. Furthermore, the report provides detailed cost analysis, supply chain. Technological innovation and advancement will further optimize the performance of the product, making it more widely used in downstream applications. Moreover, Consumer behavior analysis and market dynamics (drivers, restraints, opportunities) provides crucial information for knowing the Stem Cell Therapy market.

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Most important types of Stem Cell Therapy products covered in this report are:

Most widely used downstream fields of Stem Cell Therapy market covered in this report are:

Key Takeaways from the Global Stem Cell Therapy Market Report:

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Following Chapter Covered in the Stem Cell Therapy Market Research:

Chapter 1 mainly defines the market scope and introduces the macro overview of the industry, with an executive summary of different market segments ((by type, application, region, etc.), including the definition, market size, and trend of each market segment.

Chapter 2 provides a qualitative analysis of the current status and future trends of the market. Industry Entry Barriers, market drivers, market challenges, emerging markets, consumer preference analysis, together with the impact of the COVID-19 outbreak will all be thoroughly explained.

Chapter 3 analyzes the current competitive situation of the market by providing data regarding the players, including their sales volume and revenue with corresponding market shares, price and gross margin. In addition, information about market concentration ratio, mergers, acquisitions, and expansion plans will also be covered.

Chapter 4 focuses on the regional market, presenting detailed data (i.e., sales volume, revenue, price, gross margin) of the most representative regions and countries in the world.

Chapter 5 provides the analysis of various market segments according to product types, covering sales volume, revenue along with market share and growth rate, plus the price analysis of each type.

Chapter 6 shows the breakdown data of different applications, including the consumption and revenue with market share and growth rate, with the aim of helping the readers to take a close-up look at the downstream market.Chapter 7 provides a combination of quantitative and qualitative analyses of the market size and development trends in the next five years. The forecast information of the whole, as well as the breakdown market, offers the readers a chance to look into the future of the industry.

Chapter 8 is the analysis of the whole market industrial chain, covering key raw materials suppliers and price analysis, manufacturing cost structure analysis, alternative product analysis, also providing information on major distributors, downstream buyers, and the impact of COVID-19 pandemic.

Chapter 9 shares a list of the key players in the market, together with their basic information, product profiles, market performance (i.e., sales volume, price, revenue, gross margin), recent development, SWOT analysis, etc.

Chapter 10 is the conclusion of the report which helps the readers to sum up the main findings and points.

Chapter 11 introduces the market research methods and data sources.

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The report delivers a comprehensive study of all the segments and shares information regarding the leading regions in the market. This report also states import/export consumption, supply and demand Figures, cost, industry share, policy, price, revenue, and gross margins.

Client Focus:

Yes. As the COVID-19 and the Russia-Ukraine war are profoundly affecting the global supply chain relationship and raw material price system, we have definitely taken them into consideration throughout the research

With the aim of clearly revealing the competitive situation of the industry, we concretely analyze not only the leading enterprises that have a voice on a global scale, but also the regional small and medium-sized companies that play key roles and have plenty of potential growth.

Both Primary and Secondary data sources are being used while compiling the report.

Primary sources include extensive interviews of key opinion leaders and industry experts (such as experienced front-line staff, directors, CEOs, and marketing executives), downstream distributors, as well as end-users.

Secondary sources include the research of the annual and financial reports of the top companies, public files, new journals, etc. We also cooperate with some third-party databases.

Yes. Customized requirements of multi-dimensional, deep-level and high-quality can help our customers precisely grasp market opportunities, effortlessly confront market challenges, properly formulate market strategies and act promptly, thus to win them sufficient time and space for market competition.

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Detailed TOC of Stem Cell Therapy Market Forecast Report 2023-2028:

1 Stem Cell Therapy Market Overview

1.1 Product Overview and Scope of Stem Cell Therapy Market

1.2 Stem Cell Therapy Market Segment by Type

1.2.1 Global Stem Cell Therapy Market Sales Volume and CAGR (%) Comparison by Type (2018-2028)

1.3 Global Stem Cell Therapy Market Segment by Application

1.3.1 Stem Cell Therapy Market Consumption (Sales Volume) Comparison by Application (2018-2028)

1.4 Global Stem Cell Therapy Market, Region Wise (2018-2028)

1.5 Global Market Size of Stem Cell Therapy (2018-2028)

1.5.1 Global Stem Cell Therapy Market Revenue Status and Outlook (2018-2028)

1.5.2 Global Stem Cell Therapy Market Sales Volume Status and Outlook (2018-2028)

1.6 Global Macroeconomic Analysis

1.7 The impact of the Russia-Ukraine war on the Stem Cell Therapy Market

2 Industry Outlook

2.1 Stem Cell Therapy Industry Technology Status and Trends

2.2 Industry Entry Barriers

2.2.1 Analysis of Financial Barriers

2.2.2 Analysis of Technical Barriers

2.2.3 Analysis of Talent Barriers

2.2.4 Analysis of Brand Barrier

2.3 Stem Cell Therapy Market Drivers Analysis

2.4 Stem Cell Therapy Market Challenges Analysis

2.5 Emerging Market Trends

2.6 Consumer Preference Analysis

2.7 Stem Cell Therapy Industry Development Trends under COVID-19 Outbreak

2.7.1 Global COVID-19 Status Overview

2.7.2 Influence of COVID-19 Outbreak on Stem Cell Therapy Industry Development

3 Global Stem Cell Therapy Market Landscape by Player

3.1 Global Stem Cell Therapy Sales Volume and Share by Player (2018-2023)

3.2 Global Stem Cell Therapy Revenue and Market Share by Player (2018-2023)

3.3 Global Stem Cell Therapy Average Price by Player (2018-2023)

3.4 Global Stem Cell Therapy Gross Margin by Player (2018-2023)

3.5 Stem Cell Therapy Market Competitive Situation and Trends

3.5.1 Stem Cell Therapy Market Concentration Rate

3.5.2 Stem Cell Therapy Market Share of Top 3 and Top 6 Players

3.5.3 Mergers and Acquisitions, Expansion

4 Global Stem Cell Therapy Sales Volume and Revenue Region Wise (2018-2023)

4.1 Global Stem Cell Therapy Sales Volume and Market Share, Region Wise (2018-2023)

4.2 Global Stem Cell Therapy Revenue and Market Share, Region Wise (2018-2023)

4.3 Global Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.4 United States Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.4.1 United States Stem Cell Therapy Market Under COVID-19

4.5 Europe Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.5.1 Europe Stem Cell Therapy Market Under COVID-19

4.6 China Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.6.1 China Stem Cell Therapy Market Under COVID-19

4.7 Japan Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.7.1 Japan Stem Cell Therapy Market Under COVID-19

4.8 India Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.8.1 India Stem Cell Therapy Market Under COVID-19

4.9 Southeast Asia Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.9.1 Southeast Asia Stem Cell Therapy Market Under COVID-19

4.10 Latin America Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.10.1 Latin America Stem Cell Therapy Market Under COVID-19

4.11 Middle East and Africa Stem Cell Therapy Sales Volume, Revenue, Price and Gross Margin (2018-2023)

4.11.1 Middle East and Africa Stem Cell Therapy Market Under COVID-19

5 Global Stem Cell Therapy Sales Volume, Revenue, Price Trend by Type

5.1 Global Stem Cell Therapy Sales Volume and Market Share by Type (2018-2023)

5.2 Global Stem Cell Therapy Revenue and Market Share by Type (2018-2023)

5.3 Global Stem Cell Therapy Price by Type (2018-2023)

5.4 Global Stem Cell Therapy Sales Volume, Revenue and Growth Rate by Type (2018-2023)

6 Global Stem Cell Therapy Market Analysis by Application

6.1 Global Stem Cell Therapy Consumption and Market Share by Application (2018-2023)

6.2 Global Stem Cell Therapy Consumption Revenue and Market Share by Application (2018-2023)

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2023 Stem Cell Therapy Market Key Trends: Dynamics Shaping the ... - Digital Journal

Arthritis: The Latest on Joint Replacement, Repair, and New … – Healthline

For the millions of Americans who live with arthritis (inflammation of the joints that can affect one or multiple joints), the condition can significantly impact ones quality of life.

For those who have osteoarthritis, the degenerative disease is caused by the regular mechanical wear-and-tear on the joints over time. Those who have rheumatoid arthritis find the autoimmune disease causing pain and inflammation throughout their body, with their own immune systems attacking the tissue lining in their joints.

Arthritis can affect ones ability to get around easily, perform common everyday tasks, and simply live in comfort without inflammation and pain.

About 24% of all adults in the United States have arthritis, which is a leading cause of work disability, leading to a total annual cost of wages lost and escalating medical bills totaling $303.5 billion, according to the Centers for Disease Control and Prevention (CDC).

Many may have to turn to joint repair and replacement procedures as a way to treat their arthritis. Its so common that about 790,000 total knee replacements and 450,000 hip replacements are performed each year in the U.S., a number that will only keep growing as the nations population ages, reports the American College of Rheumatology.

To address this need, a range of technological innovations and advancements in treatment have emerged in recent years to create more long-lasting, effective treatments to repair joints affected by arthritis.

Healthline spoke with experts about the latest advancements from growing new cartilage cells to using injectables to spur more efficient healing in joint repair for arthritis, and whats next as we look to the future of the field.

When asked to assess the overall state of the field of joint repair and replacement, Dr. Susan Goodman, an attending rheumatologist at the Hospital for Special Surgery in New York, said she believes we are looking at a future with no need for joint replacements.

But to get to that point, she told Healthline that there are several significant hurdles ahead.

For a condition such as rheumatoid arthritis [RA], the problem of joint damage develops from the unchecked inflammation that erodes cartilage. For patients with RA, it is critical to control the inflammatory disease so that the new or engineered cartilage doesnt get damaged in the same way, Goodman said. For the time being, since artificial/engineered tissue has only been used in small areas of the damaged joint, it would not be a solution for patients with inflammatory arthritis who have abnormalities in their entire joint.

When it comes to joints like the knee, which are very susceptible to mechanical forces of weight and impact, a condition like obesity will also lead to damage in the engineered joint.

Dr. Kristofer Jones, a board-certified, fellowship-trained orthopedic surgeon who specializes in sports-related musculoskeletal injuries of the knee, shoulder, and elbow, told Healthline that we are ahead of where we were 10 to 15 years ago and a lot of new research has come out along with a lot of new products that look at alternative ways to resurface new cartilage, with either a patients own cells or using allograft tissue, or transplanted tissue between patients.

The research shows these new tissue types are certainly durable and provide patients with long-lasting pain relief, but the issue is the progression of joint degeneration in other areas of the knee, Jones added.

He said its not uncommon to have performed a cartilage transplant procedure to address one part of the knee and then two or three years later see the same patient experience degeneration in other areas, with new symptoms.

We are good right now at resurfacing small-to-medium size lesions with the durable tissue, but we havent quite figured out how to turn off the button that has started in some of these patients where you are looking at progressive joint degeneration in other areas, Jones explained.

From his perspective, Dr. Sid Padia, a specialist in vascular and interventional radiology at UCLA Medical Center, told Healthline that, in general, there really has been no significant impact or change in the standard of care for patients with joint disease.

Theres been no seismic shift reorienting how we view the treatment of people with joint disease, but there have been several promising and new therapies that have shown potential benefits in the treatment of various joint diseases. That being said, much of these potential new therapies currently being studied have not come to fruition with respect to long term clinical benefit, he said.

Many of the new minimally invasive therapies have shown in studies or have shown short term benefit and thats because these studies have not assessed long-term outcomes and have not compared it [the given procedure] to a control group, so its hard for the medical community to really accept these new treatment options, Padia added. So, I think there is a tremendous opportunity for a breakthrough treatment simply because in many of these patients, the treatment options are still quite limited.

Jones said one of the current advancements that stands out the most to him is the use of biologic injectables, or orthobiologics.

These are injectable substances used by orthopaedic surgeons to help your injuries heal more quickly. They can be used for tendons, ligaments, and broken bones, for example, and are derived from substances that naturally occur in your body, according to the American Academy of Orthopaedic Surgeons.

Jones cited injectable therapies like those using platelet-rich plasma (PRP), where the plasma is injected right into a tissue, and bone marrow aspirate concentrate, which uses bone marrow cells, as two examples.

PRP, bone marrow aspirate concentrate, amniotic suspension allograft injections these are all things that we are studying to determine how we can best utilize them to treat patients who have knee pain from arthritis, he said.

Jones explained that many of these injectable therapies that are being developed are to augment surgical procedures to move the healing process along and also better create more favorable cell homeostasis so further joint degeneration doesnt happen.

He said we can expect to see a lot more of these kinds of injections available in the coming years to treat symptomatic knee pain. Jones cited a Phase III trial for which he is the principal investigator at UCLA.

At UCLA we have this Phase III FDA trial that is looking at one of these products, the trial ended and we are currently crunching the numbers to look at the data to see if there are no adverse patient events, he said.

At UCLA, Padia has been working on an alternative to knee replacement that could offer pain relief to those individuals who might not be candidates for surgery.

You might not qualify for knee surgery due to a medical complication that puts you at high risk or if you are at advanced age, for instance. Younger people might also delay surgery due to the fact theyll ultimately need another knee replacement surgery or procedure within the next 20 years.

The procedure is called genicular artery embolization. Its a minimally invasive procedure during which particles that are smaller than grains of sand are injected by way of a small catheter into enlarged knee arteries. This only takes two hours to perform and you can head home the same day and return to regular physical activities later that day.

Weve published our results, done randomized trials that show genicular artery embolization in the knee can lead to reduction in pain and weve adapted this procedure for people with tennis elbow, which is something fairly common in people who play racket sports, Padia explained. Given the increase in the use of pickleball, we are starting to see a lot more people with tennis elbow.

He added that treating something like tennis elbow is fairly limited and surgical correction is rarely done. As a result, steroid injections are often the mainstay treatment for this condition, offering a short-term benefit.

People are often left with no other option than to quit their physical activity, so weve developed this procedure for tennis elbow and its had a very promising effect in people, he said.

A 2022 study in the journal Advanced Functional Materials highlights research out of Duke University that shed light on what was described as the first synthetic gel-based substitute for cartilage.

The researchers behind this gel say it can be pulled and pressed with more force and weight than naturally occurring cartilage. This substance is also three times more resistant to the regular wear and tear that often fuels osteoarthritis and joint pain.

A company called Sparta Biomedical is developing this hydrogel product and testing them on sheep, with human clinical trials expected to start this year, according to a press release.

To put in perspective how powerful this material is, natural cartilage can handle 5,800 to 8,500 pounds per inch of tugging and squishing before hitting its breaking point. The hydrogel is reportedly 26% stronger than natural cartilage in suspension and 66% stronger in compression, reads the release.

When asked what is exciting to him in the field outside of surgery right now, Jones pointed to synthetic forms of cartilage that are as durable if not more durable than the real thing.

We are a little further away from seeing that being used clinically, the issue is trying to figure out how to get those different synthetic tissue types to adhere to bone and be durable for the long term, Jones explained. That may be something that down the line is a possible alternative to traditional joint replacement. The whole point of all of this is to preserve your joint and the natural feel for it.

In late 2022, research from The Forsyth Institute was published in the journal Science Advances, pointing to a potential mechanism for generating new cartilage cells.

The goal of this study was to figure out how to regenerate cartilage. We wanted to determine how to control cell fate, to cause the somatic cell to become cartilage instead of bone, Dr. Takamitsu Maruyama of Forsyth said in a release.

The research contributes to a growing body of research that suggests future of joint repair to treat arthritis might be at the cellular level.

I think the greatest potential [for the future] are the use of stem cells, Padia said. The use of stem cells is, number one, a relatively straightforward minimally invasive procedure. It does have the potential to have significant impact on people with joint diseases. The key is there needs to be an appropriate and accurate way to select the ideal patients who benefit from a stem cell treatment, there needs to be comparative studies ideally with a placebo studies conducted with adequate amount of time for long-term results.

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Arthritis: The Latest on Joint Replacement, Repair, and New ... - Healthline

Global Cell And Gene Therapy Market Size To Grow At A CAGR Of 19.02% During The Forecast Period Of 2023-2031 – openPR

The 'Global Cell and Gene Therapy Market Size, Share, Trends, Growth, Analysis, Key Players, Report and Forecast 2023-2031' by Expert Market Research gives an extensive outlook of the global cell and gene therapy market, assessing the market on the basis of its segments like therapeutic class, type, product type, end-user, and major regions.

The key highlights of the report include:

Market Overview (2016-2031)

Forecast CAGR (2023-2031): 19.02%

The global cell and gene therapy market is expected to grow at a significant rate during the forecast period. The market growth is attributed to the increasing prevalence of chronic diseases such as cancer, genetic disorders, and others, rising investments in research and development, and increasing adoption of advanced technologies in healthcare.

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North America is expected to dominate the global cell and gene therapy market owing to the presence of a large number of key players in the region, advanced healthcare infrastructure, and increasing government funding for research and development. The Asia Pacific region is expected to grow at the highest rate during the forecast period due to the increasing adoption of advanced technologies in healthcare, rising healthcare expenditure, and the presence of a large patient population.

Europe is expected to hold a significant share of the global cell and gene therapy market due to the presence of a large number of key players in the region and increasing government funding for research and development.

Overall, the global cell and gene therapy market is expected to witness a significant growth rate during the forecast period due to the increasing prevalence of chronic diseases, rising investments in research and development, and increasing adoption of advanced technologies in healthcare.

Cell and Gene Therapy Industry Definition and Major Segments

Cell and gene therapy refers to the use of living cells and genetic material to treat or prevent diseases. This can include the use of stem cells to regenerate damaged tissue, the use of genetically modified cells to attack cancer cells, and the delivery of therapeutic genes to correct genetic disorders. Cell and gene therapy is a rapidly growing field of medicine that holds great promise for the treatment of a wide range of diseases.

Market Breakup by Therapeutic Class

Rare DiseasesOncologyHaematologyCardiologyOphthalmologyNeurologyOthers

Market Breakup by Type

Cell Therapy TypesAutologous Cell TherapyAutogenic Cell TherapyEx-vivo Cell TherapyIn-vivo Cell TherapyGene Therapy TypesSomatic Cell Gene TherapyGermline Gene TherapyEx-vivo Gene TherapyIn-vivo Gene Therapy

Market Breakup by Product Type

YescartaProvengeLuxturaKymriahImlygicGintuitMACILavivGendicineOncorineNeovasculgenStrimvelisInvossaZolgenesmaTecartusLisocelZyntelegoOthers

Market Breakup by End User

HospitalsAmbulatory Surgical CentresWound Care CentresCancer Care CentresOthers

Market Breakup by Region

North AmericaEuropeAsia PacificMiddle East and AfricaLatin America

Read Full Report with Table of Contents- https://www.expertmarketresearch.com/reports/cell-and-gene-therapy-cgt-market

Cell and Gene Therapy Market Trends

The growth of the cell and gene therapy market is driven by advances in technology and research, as well as increasing investment in the field. The increasing understanding of genetic disorders and the development of new genetic engineering techniques, such as CRISPR, have led to the development of more targeted and effective therapies.

The use of stem cells in regenerative medicine has the potential to revolutionize the treatment of a wide range of diseases. The growing prevalence of chronic diseases, such as cancer and genetic disorders, is also driving demand for cell and gene therapies. The increasing ageing population, coupled with the high costs of traditional therapies, is also expected to drive the market.

Furthermore, increasing government funding and collaborations between pharmaceutical companies and academic institutions are also driving the growth of the market. The increasing number of clinical trials and FDA approvals for cell and gene therapy products is also expected to drive market growth. However, the high cost of these therapies and the lack of reimbursement options may pose a challenge to market growth.

Key Market Players

The major players in the cell and gene therapy market report are:

Amgen, Inc.Bluebird Bio, Inc.Castle Creek Pharmaceutical HoldingsKite Pharma, Inc.Novartis AGOrchard Therapeutics plc.Pfizer, Inc.Spark Therapeutics, Inc.Vericel CorporationDendreon Pharmaceuticals LLC.Human Stem Cells InstituteDendreon Pharmaceuticals LLC.Kolon Tissuegene Inc.Organogenesis Holdings Inc.Renova Therapeutics.Others

The report studies the latest updates in the market, along with their impact across the market. It also analyses the market demand, together with its price and demand indicators. The report also tracks the market on the bases of SWOT and Porter's Five Forces Models.

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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.

This release was published on openPR.

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Global Cell And Gene Therapy Market Size To Grow At A CAGR Of 19.02% During The Forecast Period Of 2023-2031 - openPR

Association between birth by caesarian section and anxiety, self … – BMC Psychiatry

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Structural basis of spike RBM-specific human antibodies … – Nature.com

Convalescent and uninfected human blood samples

Volunteers aged 23 to 93 with a history of convalescent COVID-19 were enrolled from April 2020 to January 2021. Blood samples were collected on the day or one day before discharging from the hospital after symptom resolution. Duration is the time between PCR positive and blood sample collection. All blood samples used in this study were collected before taking any SARS-CoV-2 vaccination. Uninfected healthy volunteers aged 36 to 62 who do not have severe immunological symptoms such as immunodeficiency, autoimmune, and allergic diseases were enrolled, and we confirmed uninfected/unvaccinated donors by their clinical history and ELISA titer. Detailed information on the cohort is in Supplementary Table1. PBMCs and plasma samples were isolated by density gradient centrifugation with Ficoll-Paque PLUS (GE Healthcare) and stored at 80C until use. The study was approved by the Ethical Committee for Epidemiology of Hiroshima University (E-2011) for studies involving humans. Informed consent was obtained from all subjects involved in the study.

For single-cell sorting, PBMCs were treated with FcX blocking antibodies (BioLegend, #4422302) to reduce non-specific labeling of the cells. PBMCs were stained with S-trimer-Strep-tag, CD19-APC-Cy7 (BioLegend, #302217), and IgD-FITC (BioLegend, #348206) for 20min on ice. After washing, cells were stained with Strep-Tactin XT-DY649 (IBA) for 20min on ice. The cells were resuspended in FACS buffer (PBS containing 1% FCS, 1mM EDTA, and 0.05% NaN3) supplemented with 0.2g/ml propidium iodide (PI) to exclude dead cells. Cell sorting was performed on Special Order System BD FACSAria II (BD Biosciences) to isolate S-trimer+ CD19+ IgD cells from the PI live cell gate. Cells were directly sorted into a 96-well PCR plate. Plates containing single-cells were stored at 80C until proceeding to RT-PCR. Flow cytometric data were acquired on BD LSRFortessa (BD Biosciences) or CytoFLEX S (Beckman Coulter). Flow cytometric data were analyzed using BD FACSDiva (v8.0.2, BD Biosciences), CytExpert software (v2.4, Beckman Coulter), or FlowJo software (v10.8.1, BD Biosciences).

Single-cell sorted PCR plates were added to each well by 2l of pre-RT-PCR mix containing the custom reverse primers. After heating at 65C for 5min, plates were immediately cooled on ice. 2l of the pre-RT-PCR2 (PrimeScript II Reverse Transcriptase, Takara Bio) mix was added to each well. For RT reaction, samples were incubated at 45C for 40min followed by heating at 72C for 15min, then cooled on ice. For PCR amplification of full-length immunoglobulin heavy and light chain genes, PrimeSTAR DNA polymerase (Takara Bio) and custom primers were used. For first PCR, the initial denaturation at 98C for 1min was followed by 25 cycles of sequential reaction of 98C for 10s, 55C for 5s, and 72C for 1.5min. For second PCR, the initial denaturation at 98C for 1min was followed by 35 cycles of sequential reaction of 98C for 10s, 58C for 5s, and 72C for 1.5min. PCR fragments were assembled into a linearized pcDNA vector using NEBuilder HiFi DNA Assembly Master Mix (New England Biolabs) according to the manufacturers instructions. The pcDNA3 (Invitrogen) vectors containing an Ig light chain gene and the pcDNA4 (Invitrogen) vectors containing an Ig heavy chain gene were simultaneously transfected into Expi293 cells using Expi293 Expression System Kit (Thermo Fisher Scientific). Four days after the transfection, the culture supernatants were collected and subjected to ELISA.

For the production of recombinant S-trimer, soluble S protein (amino acids 11213), including the T4 foldon trimerization domain, a histidine tag, and a strep-tag, was cloned into the mammalian expression vector. The protein sequence was modified to remove the polybasic cleavage site (RRAR to A), and two stabilizing mutations were also introduced (K986P and V987P; wild-type numbering)34,35. The human codon-optimized nucleotide sequence encoding for the S protein of SARS-CoV-2 (GenBank: MN994467) was synthesized commercially (Eurofins Genomics). A soluble version of the S protein (amino acids 11213), including the T4 foldon trimerization domain, a histidine tag, and a strep-tag, was cloned into the mammalian expression vector pCMV. The protein sequence was modified to remove the polybasic cleavage site (RRAR to A), and two stabilizing mutations were also introduced (K986P and V987P; wild-type numbering)35. The gene encoding RBD of SARS-CoV-2, Wuhan-Hu-1, was synthesized and cloned into vector pcDNA containing a human Ig leader sequence and C-terminal 6xHis tag. RBD mutants were generated by overlap PCR using primers containing mutations. The vector was transfected into Expi293 cells and incubated at 37C for 4 days. Supernatants were purified using Capturem His-Tagged Purification kit (Takara Bio), then dialyzed by PBS buffer overnight. Protein purity was confirmed by SDS-PAGE. Protein concentration was determined spectrophotometrically at 280nm.

MaxiSorp ELISA plates (Thermo Fisher Scientific) were coated with 2g/ml purified spike RBD or trimer in 1xBBS (140mM NaCl, 172mM H3BO3, 28mM NaOH) overnight at 4C, and then blocked with blocking buffer containing 1% BSA in PBS for 1h. Antibodies diluted in Reagent Diluent (0.1% BSA, 0.05% Tween in Tris-buffered Saline) were added and incubated for 2h. HRP-conjugated antibodies were added and incubated for 2h. Wells were reacted with the TMB substrate (KPL) and the reaction was stopped using 1M HCl. The absorbance at 450nm was measured on iMark Microplate Reader (Bio-Rad) and analyzed on MPM 6 software (Bio-Rad). Antigen-specific Ig titers were determined using serial serum dilution on antigen-coated wells next to Ig-capturing antibody standard wells on the same ELISA plate.

The binding affinity of obtained antibodies to RBD was examined by the BLItz system (Sartorius Japan) using protein A-coated biosensors. 10g/ml of antibody was captured by the biosensor and equilibrated, followed by sequential binding of each concentration of RBD. For dissociation, biosensors were dipped in PBST for 900sec. Results were analyzed on BLItz Pro (v1.3.1.3, Molecular Devices).

psPAX2 (Addgene, no.12260) was a gift from Didier Trono. pCDNA3.3_CoV2_B.1.1.7 (Addgene, no.170451) for Alpha-S and pcDNA3.3-SARS2-B.1.617.2 (Addgene, no.172320) for Delta-S proteins, were gifts from David Nemazee36. pTwist-SARS-CoV-2 18 B.1.351v1 (Addgene, no.169462) for Beta-S protein was a gift from Alejandro Balazs37. Lentiviral vector, pWPI-ffLuc-P2A-EGFP for luciferase reporter assay and pTRC2puro-ACE2-P2A-TMPRSS2 for the generation of 293T cell line susceptible to SARS-CoV-2 infection was created from pWPI-IRES-Puro-Ak-ACE2-TMPRSS2, a gift from Sonja Best (Addgene, no.154987) by In-Fusion technology (Takara Bio). pcDNA3.4 expression plasmids encoding SARS-CoV-2 S proteins with human codon optimization and 19 a.a deletion of C-terminus (C-del19) from Wuhan, D614G, and Omicron were generated by assembly of PCR products, annealed oligonucleotides, or artificial synthetic gene fragments (Integrated DNA Technologies, IDT) using In-Fusion technology. For Delta plus, Kappa and Lambda variants, S proteins with only RBD, D614, and P681 mutations were created from pcDNA3.4 encoding human codon-optimized Wuhan S protein (C-del19). LentiX-293T cells (Takara Bio) and 293T cells were maintained in culture with Dulbeccos Modified Eagles Medium (DMEM) containing 10% fetal bovine serum (FBS), penicillin-streptomycin (Nacalai tesque), and 25mM HEPES (Nacalai tesque).

To generate stable 293T-ACE2.TMPRSS2 cells (293T/TRCAT), lentiviral vector VSV-G-pseudotyped lentivirus carrying ACE2 and TMPRSS2 genes were produced in LentiX-293T cells (Clontech) by transfecting with pTRC2puro-ACE2-P2A-TMPRSS2, psPAX2 (gag-pol), and pMD2G-VSV-G (envelope) using PEI-MAX (Polysciences). Packaged lentivirus was used to transduce 293T cells (Applied Biological Materials) in the presence of 5g/mL polybrene. At 72h post-infection, the resulting bulk transduced population positive for Human ACE2 expression stained by FITC-anti-ACE2 Antibody (Sinobiological) was sorted by Special Order System BD FACSAria II (BD Biosciences) and maintained in the culture medium in the presence of 2g/ml of puromycin.

Pseudoviruses bearing SARS-Cov2 S-glycoprotein and carrying a firefly luciferase (ffLuc) reporter gene were produced in LentiX-293T cells by transfecting with pWPI-ffLuc-P2A-EGFP, psPAX2, and either of S variant from Wuhan, D614G, Alpha, Beta, Delta, Delta plus, Kappa, Lambda, or Omicron using PEI-MAX (Polyscience). Pseudovirus supernatants were collected approximately 72h post-transfection and used immediately or stored at 80C. Pseudovirus titers were measured by infecting 293T/TRCAT cells for 72h before measuring luciferase activity (ONE-Glo Luciferase Assay System, Promega, Madison, WI). Pseudovirus titers were expressed as relative luminescence units per milliliter of pseudovirus supernatants (RLU/ml). For neutralization assay, pseudoviruses with titers of 14106RLU/ml were incubated with antibodies or sera for 0.5h at 37C. Pseudovirus and antibody mixtures (50l) were then inoculated with 5g/ml of polybrene onto 96-well plates that were seeded with 50l of 1104 293T/TRCAT cells/well one day before infection. Pseudovirus infectivity was scored 72h later for luciferase activity measured on ARVO X13 and 2030 Workstation (Perkin Elmer). The serum dilution or antibody concentration causing a 50% reduction of RLU compared to control (ED50 or IC50, respectively) were reported as the neutralizing antibody titers. ED50 or IC50 were calculated using a nonlinear regression curve fit on Prism (v9.0, GraphPad).

VeroE6/TMPRSS2 cells (African green monkey kidney-derived cells expressing human TMPRSS2, purchased from the Japanese Collection of Research Bioresources (JCRB) Cell Bank, JCRB1819, were maintained in DMEM containing 10% FBS and 1mg/ml G418 at 37C in 5% CO2. The virus was propagated in VeroE6/TMPRSS2 cells and the virus titer was determined by the 50% tissue culture infectious dose (TCID50) method and expressed as TCID50/ml38. The viral strains used are SARS-CoV-2/JP/Hiroshima-46059T/2020 (B.1.1, D614G, EPI_ISL_628993239), SARS-CoV-2/JP/HiroC77/2021, (AY.29, Delta, EPI_ISL_6316561), and SARS-CoV-2/JP/FH-229/2021 (BA.1.1, Omicron, EPI_ISL_11505197).

The serially diluted antibody (50l) was mixed with 100 TCID50/50l of the virus and reacted at 37C for 1h, then inoculated into VeroE6/TMPRSS2 cells to determine the minimum inhibitory concentration (MIC). Alternatively, the infectivity of the reacted antibody-virus mixture was measured by inoculating to 8 wells of a 96-well plate and observing cytopathic effects (CPE) or by the plaque assay using 10% methylcellulose to determine 50% effective dose (ED50) of the antibody. SARS-CoV-2 infection was performed in the BSL3 facility of Hiroshima University.

SARS-CoV-2 was incubated with mAb at twice the concentration of the EC50 corresponding to that viral load at 37C for 60min. After the incubation, 100l of the mixture was added to one well of a 24-well plate with confluent VeroE6/TMPRSS2 cells and incubated for 72h at 37C with 5% CO2. The supernatants were collected as an escape-mutant virus when CPE was manifested. A no-antibody-control was included to confirm the amount of test virus required.

Viral RNA was extracted from virus-infected culture medium by using Maxwell RSC Instrument (Promega, AS4500). cDNA preparation and amplification were done in accordance with protocols published by the ARTIC network (https://artic.network/ncov-2019) using V4 version of the ARTIC primer set from Integrated DNA Technologies to create tiled amplicons across the virus genome. The sequencing library was prepared using the NEB Next Ultra II DNA Library Prep Kit for Illumina (New England Biolabs, E7645). Paired-end, 300bp sequencing was performed using MiSeq (Illumina) with the MiSeq reagent kit v3 (Illumina, MS-102-3003). Consensus sequences were obtained by using the DRAGEN COVID lineage software (Illumina, ver. 3.5.6). Variant calling and annotation were performed using the Nextclade website (https://clades.nextstrain.org).

RBD from Wuhan-Hu-1, Delta, and Omicron variant and Fab fragment from NCV2SG48 with 6xHis-tag expressed in Expi293F cells (Thermo Fisher Scientific) were purified using Ni-NTA Agarose resin (QIAGEN). Fab fragment of NCV2SG53 was isolated from papain digests of the monoclonal antibody expressed in Expi293F cells using HP Protein G column (Cytiva). Purified each Fab fragments and RBD were mixed in the molar ration of 1:1.2 and incubated on ice for 1h. The mixture was loaded onto a Superdex 200 increase 10/300 GL column (Cytiva) equilibrated in 20mM Tris-HCl pH7.5, 150mM NaCl for removing the excess RBD. Fractions containing RBD and each Fab were collected and concentrated for crystallization. Chromatography was performed using NGC Chromatography Systems (BIO-RAD) and ChromLab v6 (BIO-RAD).

Crystallization was carried out by the sitting-drop vapor diffusion method at 20C. Crystals of RBD (Wuhan-Hu-1)-Fab (NCV2SG48) were grown in 2l drops containing a 1:1 (v/v) mixture of 7.5mg/ml RBD solution and 0.1M Bis-Tris pH5.5, 0.5M ammonium sulfate and 19% PEG3350. Crystals of RBD (Delta)-Fab (NCV2SG48) were grown in 2l drops containing a 1:1 (v/v) mixture of 7.5mg/ml RBD solution and 0.1M Bis-Tris pH6.5, 17.5% PEG10000, 100mM Ammonium acetate and 5% Glycerol. Crystals of RBD (Omicron BA.1)-Fab (NCV2SG48) were grown in 2l drops containing a 1:1 (v/v) mixture of 7.5mg/ml RBD solution and 0.1M Bis-Tris pH 5.5, 0.5M ammonium sulfate, 19.5% PEG 3350, 1mM EDTA and 10% glycerol. Crystals of RBD (Wuhan-Hu-1)-Fab (NCV2SG53) were grown in 2l drops containing a 1:1 (v/v) mixture of 7.5mg/ml RBD solution and 0.1M MES pH 6.0, 0.25M ammonium sulfate and 22.5% PEG3350. Crystals of RBD (Delta)-Fab (NCV2SG53) were grown in 0.6l drops containing a 1:1 (v/v) mixture of 7.5mg/ml RBD solution and 25% PEG1500. The single crystals suitable for X-ray experiments were obtained in a few weeks. X-ray diffraction data collections were performed using synchrotron radiation at SPring-8 beamline BL44XU40 in a nitrogen vapor stream at 100K. The data sets were indexed and integrated using the XDS package41, scaled, and merged using the program Aimless42 in the CCP4 program package43. The scaling statistics were shown in Table1.

Phase determinations were carried out by the molecular replacement method using the program Phaser44 in the PHENIX package45 and the program Molrep46 in the CCP4 program package with the combination of RBD structure (PDB ID:7EAM) and Fab structures (PDB ID:7CHB and 7CHP) as search models. The structure refinement was performed using the program phenix.refine47 in the PHENIX package and the program coot48 in the CCP4 program package. The final refinement statistics were shown in Table1. Interactions between RBD and Fabs were analyzed using the program PISA49 in the CCP4 program package. All figures of structures were generated by the program pymol (The PyMOL Molecular Graphics System, Version 2.4.0., Schrdinger, LLC.). Class 2a/AZD8895 (PDB ID:7L7D), Class 3a/REGN10987 (PDB ID:6XDG), Class 3b/S309 (PDB ID:7JX3), Class 4a/CR3022 (PDB ID:6ZLR), Class 4b/S2X259 (PDB ID:7M7W), and Class 5/S2H97 (PDB ID:7M7W) open data were used in Fig.3a.

The statistical analysis was performed using Prism 9.0 (GraphPad, La Jolla, CA, USA). Ordinary One-way ANOVA, Two-way ANOVA, Kruskal-Wallis test, Wilcoxon rank test, and Friedman test were used to compare data. P-value 0.05 was considered statistically significant. Statistical tests are reported in figure legends and significance is reported at p0.05. To verify reproducibility, we repeated experiments more than two times as indicated in Figure legends. Detailed information on the sample is provided in Supplementary Tables.

Further information on research design is available in theNature Portfolio Reporting Summary linked to this article.

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