Regenerative Medicine Industry Research Articles

J-TEC's efforts to industrialize regenerative medicine in Japan

Japan Tissue Engineering Co., Ltd., J-TEC, was launched in 1999 to industrialize regenerative medicine in Japan. We developed the first regenerative medicine product, JACE (autologous cultured epidermis), which received PMDA approval for treating serious burns in 2007. Then, JACC (autologous cultured cartilage), the second product, was approved in 2012 for efficacy on traumatic cartilage defects. In 2014, the Pharmaceutical Affairs Law was revised to the Pharmaceutical and Medical Device Act, and regenerative medicine products, including gene therapies, were newly classified to accelerate productization. Subsequently, Nepic (autologous cultured corneal epithelium) and Ocural (autologous cultured oral mucosal epithelium) for epithelialization of limbal stem cell deficiencies in ophthalmology were approved in 2020 and 2021, respectively. Furthermore, a new product, JACEMIN (autologous cultured epidermis maintaining melanocyte) for vitiligo treatment was approved in 2023. We have developed five products of regenerative medicine that construct human tissues to graft rather than injectable cell suspensions like drugs. To develop regenerative medicine products, it is necessary to ensure the safety of raw materials, standardize the cultivation process, examine cell characteristics on GLP tests, construct transportation methods, build GCTP facilities, and conduct clinical trials on GCP. Re-examinations of JACE for serious burns and JACC for cartilage defects were completed after 7 years of all-case postmarketing surveillance. The commercialization of these products has become a benchmark for domestic regulation and has induced the development of a regenerative medicine industry promoted by Japan.

Brief exposure to directionally-specific pulsed electromagnetic fields stimulates extracellular vesicle release and is antagonized by streptomycin: A potential regenerative medicine and food industry paradigm

Pulsing electromagnetic fields (PEMFs) have been shown to promote in vitro and in vivo myogeneses via mitohormetic survival adaptations of which secretome activation is a key component. A single 10-min exposure of donor myoblast cultures to 1.5 mT amplitude PEMFs produced a conditioned media (pCM) capable of enhancing the myogenesis of recipient cultures to a similar degree as direct magnetic exposure. Downwardly-directed magnetic fields produced greater secretome responses than upwardly-directed fields in adherent and fluid-suspended myoblasts. The suspension paradigm allowed for the rapid concentrating of secreted factors, particularly of extracellular vesicles. The brief conditioning of basal media from magnetically-stimulated myoblasts was capable of conferring myoblast survival to a greater degree than basal media supplemented with fetal bovine serum (5%). Downward-directed magnetic fields, applied directly to cells or in the form of pCM, upregulated the protein expression of TRPC channels, markers for cell cycle progression and myogenesis. Direct magnetic exposure produced mild oxidative stress, whereas pCM provision did not, providing a survival advantage on recipient cells. Streptomycin, a TRP channel antagonist, precluded the production of a myogenic pCM. We present a methodology employing a brief and non-invasive PEMF-exposure paradigm to effectively stimulate secretome production and release for commercial or clinical exploitation.

Creative regulatory practices to develop stem-cell technology: the way forward for Malaysia.

Malaysia aspires to develop regenerative medicine through stem-cell technology. It needs a regulatory system that could facilitate development and prevent unethical practices. A comparative legal analysis on the regulation of stem-cell technology, with a focus on stem-cell research in Malaysia and selected Commonwealth countries that are experienced in regulating this complex technology, demonstrates that the selected Commonwealth countries have adopted a hybrid of different regulatory mechanisms. This paper argues that Malaysia should consider adopting a similar approach to equip relevant authorities with different regulatory mechanisms that are able to promote innovation in stem-cell research activities and cultivate a successful and profitable regenerative medicine industry in the future. Such a strategic action can produce an optimal regulatory outcome and help Malaysia to realize its aspiration.

Https://www.insights.bio/cell-and-gene-therapy-insights/journal/article/2128/Scalable-upstream-process-development-for-the-suspension-based-production-of-lentiviral-vectors-for-CAR-T-cell-therapies-with-multiparallel-benchtop

Cell and gene-based therapies present a new treatment paradigm that have the potential to address unmet clinical needs. Viral vectors such as adenoviruses, adeno-associated viruses, retroviruses and lentiviruses are effective delivery systems for genetic material used in cell and gene therapies and vaccines. HEK-293 cells and derivatives are commonly used as a workhorse cell line for lentiviral vector (LVV) production for cell and gene therapy applications. Adherent production processes with these cells utilize static flask cultures, and this adherent method is quite easy to develop and perform. However, it also significantly lacks the ability for automation and scalability. Typical bioreactors based either on a rocking motion or stirred tank agitation can provide these features. Therefore, to scale up viral vector production for commercialization, adherent processes should be shifted to a suspension-based process, a significant challenge for the regenerative medicine industry. The use of a suspension adapted HEK-293 cell line and the Ambr^{^® }15 microbioreactor system can facilitate transition from adherent cultures to suspension cultures by enabling fast process optimization with the ability to screen in parallel many parameters in small volumes. As a proof-of-concept study, we established here such a transitional protocol for the cultivation of suspension adapted HEK-293T cells and the production of CD19-CAR lentivirus in small and benchtop scale stirred bioreactors.

3D Bioprinting at the Frontier of Regenerative Medicine, Pharmaceutical, and Food Industries.

3D printing technology has emerged as a key driver behind an ongoing paradigm shift in the production process of various industrial domains. The integration of 3D printing into tissue engineering, by utilizing life cells which are encapsulated in specific natural or synthetic biomaterials (e.g., hydrogels) as bioinks, is paving the way toward devising many innovating solutions for key biomedical and healthcare challenges and heralds' new frontiers in medicine, pharmaceutical, and food industries. Here, we present a synthesis of the available 3D bioprinting technology from what is found and what has been achieved in various applications and discussed the capabilities and limitations encountered in this technology.

Exosome and Biomimetic Nanoparticle Therapies for Cardiac Regenerative Medicine.

Exosomes and biomimetic nanoparticles have great potential to develop into a wide-scale therapeutic platform within the regenerative medicine industry. Exosomes, a subgroup of EVs with diameter ranging from 30-100 nm, have recently gained attention as an innovative approach for the treatment of various diseases, including heart disease. Their beneficial factors and regenerative properties can be contrasted with various cell types. Various biomimetic nanoparticles have also emerged as a unique platform in regenerative medicine. Biomimetic nanoparticles are a drug delivery platform, which have the ability to contain both biological and fabricated components to improve therapeutic efficiency and targeting. The novelty of these platforms holds promise for future clinical translation upon further investigation. In order for both exosome therapeutics and biomimetic nanoparticles to translate into large-scale clinical treatment, numerous factors must first be considered and improved. Standardization of different protocols, from exosome isolation to storage conditions, must be optimized to ensure batches are pure. Standardization is also important to ensure no variability in this process across studies, thus making it easier to interpret data across different disease models and treatments. Expansion of clinical trials incorporating both biomimetic nanoparticles and exosomes will require a standardization of fabrication and isolation techniques, as well as stricter regulations to ensure reproducibility across various studies and disease models. This review will summarize current research on exosome therapeutics and the application of biomimetic nanoparticles in cardiac regenerative medicine, as well as applications for exosome expansion and delivery on a large clinical scale.

Rapid transient expression of functional human vascular endothelial growth factor in Nicotiana benthamiana and characterization of its biological activity

Human vascular endothelial growth factor (VEGF) is a potent pro-angiogenic growth factor essential for wound healing. Due to its potential applications, many expression strategies have been developed to produce high levels of VEGF. Here, we have optimized the expression conditions for the production of recombinant VEGF in Nicotiana benthamiana by using a geminiviral vector. Four different expression constructs that differ by the location of a C- or N-terminal histidine tag and SEKDEL sequence were developed and utilized for plant transient expression. The recombinant VEGF was further purified by using affinity chromatography and confirmed by SDS-PAGE and Western blotting probed with anti-VEGF antibody. Furthermore, our results showed that the recombinant VEGF in all tested concentrations did not exhibit any cytotoxic effect on HaCaT cells and induced cell migration in vitro. These findings show that the plant-produced VEGF has the potential to be used in regenerative medicine and cosmetic industry.

3D Bioprinting: The Roller Coaster Ride to Commercialization.

Three-dimensional (3D) bioprinting as a technology is being researched and applied since 2003. It is actually several technologies (inkjet, extrusion, laser, magnetic bioprinting, etc.) under an umbrella term “3D bioprinting.” The versatility of this technology allows widespread applications in several; however, after almost 20 years of research, there is still a limited number of cases of commercialized applications. This article discusses the potential for 3D bioprinting in regenerative medicine, drug discovery, and food industry, as well as the existing cases of companies that create commercialized products and services in the aforementioned areas and even in fashion, including their go-to-market route and financing received. We also address the main barriers to creating practical applications of 3D bioprinting within each sphere the technology that is being studied for.

The new role of a science park for the current startup ecosystem in Japan：A case study on regenerative medicine industry

The new role of a science park for the current startup ecosystem in Japan：A case study on regenerative medicine industry

Efficient Expression, Purification and Functional Evaluation of Human Epidermal Growth Factor-Collagen Binding Domain Hybrid Protein on Growth Stimulation of Human Fibroblast Cells

Growth factors such as human epidermal growth factor (hEGF) have recently received high interest in regenerative medicine and pharmaceutical industries mainly due to their ability to restore tissues proliferation and improvement of their biological functions. In spite of various hEGF applications, its efficient expression in Escherichia coli could not yet reach an industrial reality mainly due to the lack of the ability of folding into the correct 3D structure because of three disulfide bonds in monomer hEGF. To address these challenges, here a fusion hEGF protein with a C terminus of collagen binding domain (CBD) along with intein protein with self-splicing property and ELP sequence was constructed by a three-step cloning procedure. This enabled us to purify recombinant hEGF without using chromatography columns. Following the confirmation of the construct by colony PCR, restriction enzymes analysis and sequencing, the 62[Formula: see text]kDa band of ELP-INTEIN-hEGF-CBD were observed on SDS-PAGE and confirmed by western blotting. Subsequently, the mitotic activity in Balb/c 3T3 cells proliferation in presence of recombinant hEGF-CBD compared with commercial hEGF using methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay which showed that our purified recombinant protein stimulates the cell proliferation similar to the commercial protein. Our strategy could be considered as a new feasible approach to produce hEGF in E. coli for pharmaceutical and clinical applications.

Advancing the robust manufacture of t-cell therapies through the application of stirred tank bioreactors

Background & Aim With over 90 active trials in the United States with many more in pre-clinical phases T-Cell immunotherapies represent a large share of the expansion we are observing in the regenerative medicine industry. While CAR-T therapies have dominated the headlines, other technologies such as TCR, TIL, Treg, CAR-NK, γδ T-Cells have shown impressive results the oncologic space and elsewhere, however a robust manufacturing process is required to realize these solutions in the clinic. A main hurdle in establishing a robust manufacturing process for these therapies is a lengthy expansion period is typically necessary to reach a relevant dose of therapeutic cells. This only becomes more critical as allogeneic cell therapies become a reality. The issues with current culture methods present with a high cost of goods, increased production timelines, and highly variant expansion rates. Methods, Results & Conclusion At Hitachi Chemical Advanced Therapeutic Solutions, we investigated stirred tank bioreactors as an alternative to standard static and rocking motion culture methods using the ambr15 microbioreactor system and confirming those results in a two liter stirred tank bioreactors. Because of the extensive history stirred tank bioreactors have in cell culture we have demonstrated that a power input of 7.5 W/m3 does not create undo shear stress on T-Cells and were able to directly translate that parameter from the 15mL scale to the 2L. This work focused on defining the specific parameters that would facilitate stirred tank bioreactors as a unique solution for the manufacture of both autologous and allogeneic T-cell therapies with high rates of mass and gas transfer and established process control methods. Given these advantages, a comparison study of T-cells expanded in the stirred tank bioreactor and batch fed static cultures resulted in a three-fold increase in total cell number with the bioreactors. Further development of stirred tank culture methods has demonstrated five population doublings over an eight-day culture period. These outcomes indicate at a T-cell expansion method that could expand the potential of the revolutionary therapies through reduced media requirements, shortened culture periods, and high yields. By further utilizing established techniques in cell culture, T-cell therapies become increasingly viable treatments for a range of indications.

A Review of Decellularized Extracellular Matrix Biomaterials for Regenerative Engineering Applications

Biomaterials are a cornerstone technology of the biomedical device, tissue engineering, and regenerative medicine industries. While traditional biomaterials are fully defined synthetics, growing evidence supports the use of extracellular matrix-based biomaterials produced through the decellularization of organs, tissues, or cell cultures. These materials are particularly advantageous as they largely retain the structure and the biochemical nature of the original tissue, properties that are often difficult to reproduce with synthetics. Indeed, there are many FDA-approved and clinically used extracellular matrix-based materials that are generated through decellularization processes. In this review, we first describe methods of decellularization used to produce these materials and their associated advantages and limitations, discuss the current use of extracellular matrix-based materials in regenerative engineering applications, describe the areas where current research is occurring, and forecast areas where impactful research may appear. The regeneration of tissues often requires a scaffold material to support and guide the cells that are performing the repair. Often, these materials are manmade and lack many of the key features present in native tissue. However, a tissue can be processed to remove its cells (a process called decellularization), leaving behind a scaffold of proteins and polysaccharides known as the extracellular matrix. These decellularized matrices are attractive scaffolds for use in regenerative medicine applications, and they are the subject of this review.

An Investigation and Analysis of Literature on Stem Cell and Regenerative Medicine Industry in China

The latest report from the National Bureau of Statistics, “China’s economic and social development in 2016”, shows that the number of people aged 60 and above is approximately 230 million (i.e., 16.7% of the population). By 2050, this age-group will increase to 400 million, accounting for more than 30% of the population. In China, this dramatic aging trend has endangered people’s health. Major disorders such as tissue and organ damage, cardiovascular and other functional organ failures, degenerative diseases, and cancer are closely related to aging. Traditional treatments such as drug therapy and surgery often have little effect on such diseases, and fail to meet the growing medical needs of this age-group. Stem cell-based regenerative medicine is expected to become the third treatment option after drug therapy and surgery. With the increasing financial supports and investments in China recent years, a series of important progress have been made to stem cells and regenerative medicine. In this issue, we investigated the stem cell and regenerative medicine industry in the world and china, such as research literature and research institutions distribution.

Corporate profile: RoosterBio, Inc.

RoosterBio, Inc. (MD, USA) is a privately held stem cell tools and technology company focused on accelerating the development of a sustainable regenerative medicine industry, one customer at a time. RoosterBio's products are high-volume and well-characterized adult human mesenchymal stem/stromal cells (hMSCs) paired with highly engineered media systems. RoosterBio has aimed to simplify and standardize how stem cells are purchased, expanded and used in the development of regenerative medicine products. To this end, RoosterBio supplies off-the-shelf cGMP hMSC working cell banks with bioprocess media that mimic the format and formulation of the research grade counterparts, radically simplifying and shortening product development and clinical translation. RoosterBio's focus is to offer innovative products that help usher in a new era of productivity and standardization into the field, with a passion directed towards empowering life-saving cures to be discovered in regenerative medicine.

Challenging the Problem of ‘Fit’: Advancing the Regenerative Medicine Industries in the United States, Britain and Japan

This article follows the evolution of biopharmaceutical firms as they bore great uncertainty and risks in their endeavours to commercialise new therapies – through the regenerative medicine industry in the United States, Britain and Japan. Despite its beginnings in the 1970s, regenerative medicines have yet to become a widely accepted form of medicine. A large part of the problem lay with the lack of ‘fit’ with the broader health context. This article illustrates how the trajectory of the sector was shaped, not only by the nature of the technology, but also by the complex contexts in which firms were embedded.

Impedance-based cellular assays for regenerative medicine.

Therapies based on regenerative techniques have the potential to radically improve healthcare in the coming years. As a result, there is an emerging need for non-destructive and label-free technologies to assess the quality of engineered tissues and cell-based products prior to their use in the clinic. In parallel, the emerging regenerative medicine industry that aims to produce stem cells and their progeny on a large scale will benefit from moving away from existing destructive biochemical assays towards data-driven automation and control at the industrial scale. Impedance-based cellular assays (IBCA) have emerged as an alternative approach to study stem-cell properties and cumulative studies, reviewed here, have shown their potential to monitor stem-cell renewal, differentiation and maturation. They offer a novel method to non-destructively assess and quality-control stem-cell cultures. In addition, when combined with in vitro disease models they provide complementary insights as label-free phenotypic assays. IBCA provide quantitative and very sensitive results that can easily be automated and up-scaled in multi-well format. When facing the emerging challenge of real-time monitoring of three-dimensional cell culture dielectric spectroscopy and electrical impedance tomography represent viable alternatives to two-dimensional impedance sensing.This article is part of the theme issue 'Designer human tissue: coming to a lab near you'.

Thermal Treatment of 3D-printed Titanium Alloy

In metals processing, 3D printing is a relatively new technology. It brings many advantages into production thanks to its additive principle on which it is based. One of the intended applications of 3D printing is especially regenerative medicine and aerospace industry that require products of very complex shapes. In these domains, titanium along with its alloys belongs among the most frequently used materials. When printing a Ti6Al4V alloy, very high cooling rates during the successive laser melting of an initial powder material result in high internal stresses. These stresses are followed with several problems, such as low material plasticity, possible cracking of built products, deformations of thin parts and similarly. Therefore, after the 3D printing process itself, a thermal treatment is applied to relief the stresses. The object of this study is to show the influence of atmosphere in thermal treatment process on the quality of final parts. The results show that oxygen absence is essential in terms of material plasticity.

Classification of C2C12 cells at differentiation by convolutional neural network of deep learning using phase contrast images

In the field of regenerative medicine, tremendous numbers of cells are necessary for tissue/organ regeneration. Today automatic cell-culturing system has been developed. The next step is constructing a non-invasive method to monitor the conditions of cells automatically. As an image analysis method, convolutional neural network (CNN), one of the deep learning method, is approaching human recognition level. We constructed and applied the CNN algorithm for automatic cellular differentiation recognition of myogenic C2C12 cell line. Phase-contrast images of cultured C2C12 are prepared as input dataset. In differentiation process from myoblasts to myotubes, cellular morphology changes from round shape to elongated tubular shape due to fusion of the cells. CNN abstract the features of the shape of the cells and classify the cells depending on the culturing days from when differentiation is induced. Changes in cellular shape depending on the number of days of culture (Day 0, Day 3, Day 6) are classified with 91.3% accuracy. Image analysis with CNN has a potential to realize regenerative medicine industry.

110 - Manufacturing solutions for mesenchymal stromal/stem cell therapy expansion

There is a growing need within the regenerative medicine industry for closed cell culture systems able to support scalable manufacture of adherent cell therapies. Single-use stirred tank bioreactor systems are such a platform, having been proven effective for large scale production of allogenic microcarrier-based cell therapies. However, optimization of the bioreactor culture process is essential to maximize cell yield while maintaining cell function. We will review a mesenchymal stem/stromal cell (MSC) case study using qualified media, serum alternatives, and bioreactor process control strategies to achieve increased growth. In this study, we present a method to determine optimal bioreactor process parameters, such as agitation set-point, pH set-point, and microcarrier concentration in the Mobius® 3L single-use bioreactor for MSC expansion on microcarriers. Specifically, we have utilized Zwietering's equation for suspension of solids in stirred tanks to determine a theoretical agitation operating range for microcarrier cultures, and Kolmogorov's theory of turbulent eddy lengths to define upper agitation limits. Results from this study suggest it is critical to balance minimizing shear and maximizing cell access to nutrients, DO, and cell-to-microcarrier attachment and transfer to achieve optimal MSC growth. Specifically this work indicates an agitation rate creating a slight gradient of microcarriers, produces higher growth compared to a more traditional homogenous distribution. Additionally, MSC culture performance was found to be negatively impacted by pH values outside the range of 7.4–7.6 tested in this study. Start to finish solutions for expansion, including high quality reagents and process control strategies, are key to enabling technologies for success in commercializing cell therapies.

CCRM: cultivating a culture of cooperation to advance the global regenerative medicine industry.

Launched in June 2011, CCRM is a unique, Canadian, not-for-profit group that is solely focused on developing and commercializing regenerative medicine, cell and gene therapy technologies. Its mission is to generate sustainable health and economic benefits through global collaborations, and its vision is to be the preferred destination for the best people and companies, technologies, clinical trials and investments in cell and gene therapies, and regenerative medicine.