Regenerative Medicine Products Research Articles

Automation, Monitoring, and Standardization of Cell Product Manufacturing.

Although regenerative medicine products are at the forefront of scientific research, technological innovation, and clinical translation, their reproducibility and large-scale production are compromised by automation, monitoring, and standardization issues. To overcome these limitations, new technologies at software (e.g., algorithms and artificial intelligence models, combined with imaging software and machine learning techniques) and hardware (e.g., automated liquid handling, automated cell expansion bioreactor systems, automated colony-forming unit counting and characterization units, and scalable cell culture plates) level are under intense investigation. Automation, monitoring and standardization should be considered at the early stages of the developmental cycle of cell products to deliver more robust and effective therapies and treatment plans to the bedside, reducing healthcare expenditure and improving services and patient care.

Marine Algae Polysaccharides as Basis for Wound Dressings, Drug Delivery, and Tissue Engineering: A Review

The present review considers the physicochemical and biological properties of polysaccharides (PS) from brown, red, and green algae (alginates, fucoidans, carrageenans, and ulvans) used in the latest technologies of regenerative medicine (tissue engineering, modulation of the drug delivery system, and the design of wound dressing materials). Information on various types of modern biodegradable and biocompatible PS-based wound dressings (membranes, foams, hydrogels, nanofibers, and sponges) is provided; the results of experimental and clinical trials of some dressing materials in the treatment of wounds of various origins are analyzed. Special attention is paid to the ability of PS to form hydrogels, as hydrogel dressings meet the basic requirements set out for a perfect wound dressing. The current trends in the development of new-generation PS-based materials for designing drug delivery systems and various tissue-engineering scaffolds, which makes it possible to create human-specific tissues and develop target-oriented and personalized regenerative medicine products, are also discussed.

Phoenix: A Portable, Battery-Powered, and Environmentally Controlled Platform for Long-Distance Transportation of Live-Cell Cultures.

Despite the advent of advanced therapy medicinal products (ATMPs) in regenerative medicine, gene therapy, cell therapies, tissue engineering, and immunotherapy, the availability of treatment is limited to patients close to state-of-the-art facilities. The SCORPIO-V Division of HNu Photonics has developed the Phoenix-Live Cell TransportTM, a battery-operated mobile incubator designed to facilitate long-distance transportation of living cell cultures from GMP facilities to remote areas for increased patient accessibility to ATMPs. This work demonstrates that PhoenixTM (patent pending) is a superior mechanism for transporting living cells compared to the standard method of shipping frozen cells on dry ice (−80°C) or in liquid nitrogen (−150°C), which are destructive to the biology as well as a time consuming process. Thus, Phoenix will address a significant market need within the burgeoning ATMP industry. SH-SY5Y neuroblastoma cells were cultured in a stationary Phoenix for up to 5 days to assess cell viability and proliferation. The results show there is no significant difference in cell proliferation (∼5X growth on day 5) or viability (>90% viability on all days) when cultured in PhoenixTM and compared to a standard 5% CO2 incubator. Similarly, SH-SY5Y cells were evaluated following ground (1–3 days) and air (30 min) shipments to understand the impact of transit vibrations on the cell cultures. The results indicate that there is no significant difference in SH-SY5Y cell proliferation (∼2X growth on day 3) or viability (>90% viability for all samples) when the cells are subjected to the vibrations of ground and air transportation when compared to control samples in a standard, stationary 5% CO2 incubator. Furthermore, the temperature, pressure, humidity, and accelerometer sensors log data during culture shipment to ensure that the sensitive ATMPs are handled with the appropriate care during transportation. The PhoenixTM technology innovation will significantly increase the accessibility, reproducibility, and quality-controlled transport of living ATMPs to benefit the widespread commercialization of ATMPs globally. These results demonstrate that PhoenixTM can transport sensitive cell lines with the same care as traditional culture techniques in a stationary CO2 incubator with higher yield, less time and labor, and greater quality control than frozen samples.

Advanced therapy medicinal product manufacturing under the hospital exemption and other exemption pathways in seven European Union countries

Background aimsAs part of the advanced therapy medicinal product (ATMP) regulation, the hospital exemption (HE) was enacted to accommodate manufacturing of custom-made ATMPs for treatment purposes in the European Union (EU). However, how the HE pathway has been used in practice is largely unknown. MethodsUsing a survey and interviews, we provide the product characteristics, scale and motivation for ATMP manufacturing under HE and other, non-ATMP-specific exemption pathways in seven European countries. ResultsResults show that ATMPs were manufactured under HE by public facilities located in Finland, Germany, Italy and the Netherlands, which enabled availability of a modest number of ATMPs (n = 12) between 2009 and 2017. These ATMPs were shown to have close proximity to clinical practice, and manufacturing was primarily motivated by clinical needs and clinical experience. Public facilities used HE when patients could not obtain treatment in ongoing or future trials. Regulatory aspects motivated (Finland, Italy, the Netherlands) or limited (Belgium, Germany) HE utilization, whereas financial resources generally limited HE utilization by public facilities. Public facilities manufactured other ATMPs (n = 11) under named patient use (NPU) between 2015 and 2017 and used NPU in a similar fashion as HE. The scale of manufacturing under HE over 9 years was shown to be rather limited in comparison to manufacturing under NPU over 3 years. In Germany, ATMPs were mainly manufactured by facilities of private companies under HE. ConclusionsThe HE enables availability of ATMPs with close proximity to clinical practice. Yet in some countries, HE provisions limit utilization, whereas commercial developments could be undermined by private HE licenses in Germany. Transparency through a public EU-wide registry and guidance for distinguishing between ATMPs that are or are not commercially viable as well as public-private engagements are needed to optimize the use of the HE pathway and regulatory pathways for commercial development in a complementary fashion.

Governmental Regulations and Increasing Food and Drug Administration Oversight of Regenerative Medicine Products: What’s New in 2020?

The United States Food and Drug Administration (FDA) is responsible for protecting and promoting public health through rules and regulations. Over the past few years, the field of regenerative medicine and cell therapy have garnered significant interest, and this evolving new biology is changing fast and challenging regulatory bodies. The FDA has published a series of guidance documents outlining steps to protect consumers against potentially dangerous and unproven treatments. The agency has offered a grace period for “stem cell clinics” until November 2020 to come into compliance by obtaining Investigational New Drug applications and working to secure premarket approval of their products. With the documentation of hundreds of “stem cell clinics,” the FDA needs to enforce the adherence to their outlined standards to protect patients. The aim of this review was to provide an overview of these FDA regulations and some current issues within the industry. The purpose is to educate and inform the musculoskeletal community about the current government regulations of this new expanding biology. Level of EvidenceLevel V, expert opinion.

Development of process analytical technology to monitor the cell secretome during differentiation of pluripotent stem cells

Background & Aim Induced pluripotent stem cells (iPSC) offers a renewable source for reliably and robustly generating off-the-shelf cell products for regenerative medicine. A number of differentiation protocols have been developed and PSC-derived cells have been used in a variety of animal models to treat disease and progressed through to clinical trials in Asia and more recently the United States. However, the manufacturing processes for expansion and differentiation of iPSC cells remain reliant on poorly scalable technologies with limited in process monitoring despite typically > 14 days of culture and manipulation required; increasing the risks and costs of batch manufacturing failure. The industrialisation of pluripotent stem cell-derived cell therapies requires the development of process analytical technologies (PAT) for the monitoring and control of large-scale cell differentiation bioprocesses. The extracellular microenvironment provides fertile hunting ground to determine the impact of processing conditions on process outcome with the myriad of molecules consumed and produced by cells during differentiation alluding to cell state and potential cell fate. PAT that targets the spent media enables control of critical processing parameters to maintain product critical quality attributes without requiring cell-based analysis to be conducted. Methods, Results & Conclusion Using two exemplar differentiation processes, going from induced pluripotent stem cells to, either, definitive endoderm or cardiomyocytes, we have characterised the cell secretome with respect to metabolites and secreted proteins, and performed multi-parametric analysis to determine potential biomarkers for monitoring future large-scale differentiation processes using cost- and time-effective PAT alternatives.

Integration of nano- and biotechnology for beta-cell and islet transplantation in type-1 diabetes treatment.

Regenerative medicine using human or porcine β‐cells or islets has an excellent potential to become a clinically relevant method for the treatment of type‐1 diabetes. High‐resolution imaging of the function and faith of transplanted porcine pancreatic islets and human stem cell–derived beta cells in large animals and patients for testing advanced therapy medicinal products (ATMPs) is a currently unmet need for pre‐clinical/clinical trials. The iNanoBIT EU H2020 project is developing novel highly sensitive nanotechnology‐based imaging approaches allowing for monitoring of survival, engraftment, proliferation, function and whole‐body distribution of the cellular transplants in a porcine diabetes model with excellent translational potential to humans. We develop and validate the application of single‐photon emission computed tomography (SPECT) and optoacoustic imaging technologies in a transgenic insulin‐deficient pig model to observe transplanted porcine xeno‐islets and in vitro differentiated human beta cells. We are progressing in generating new transgenic reporter pigs and human‐induced pluripotent cell (iPSC) lines for optoacoustic imaging and testing them in transplantable bioartificial islet devices. Novel multifunctional nanoparticles have been generated and are being tested for nuclear imaging of islets and beta cells using a new, high‐resolution SPECT imaging device. Overall, the combined multidisciplinary expertise of the project partners allows progress towards creating much needed technological toolboxes for the xenotransplantation and ATMP field, and thus reinforces the European healthcare supply chain for regenerative medicinal products.

The regulation of cannabis-based medicinal products across jurisdictions

The regulation of cannabis-based medicinal products across jurisdictions

Gene therapy regulation: could in-body editing fall through the net?

Somatic gene therapies may be authorised for marketing in the EU under the advanced therapy medicinal product regulation. These therapeutic compounds are sufficiently novel and complex in their potential effects to require specialist evaluation. However, the current definition of gene therapy medicinal products (‘GTMP’) risks excluding molecules which are not manufactured through techniques involving recombination. We consider the way, in which the ‘recombinant nucleic acid’ aspect of the GTMP definition is challenged by developments in gene-editing technology, and why a broader scope of GTMP regulation may be desirable.

Regulated, reliable, and reputable: Protect patients with uniform standards for stem cell treatments.

The promise of cell and gene therapies is being realized as new products emerge to treat diseases once considered intractable. These treatments are emerging amidst reports of patients being injured by unproven “stem cell” interventions. At this juncture, it is vital to be supporting the continued development of promising regenerative medicine products while protecting patients from the risks posed by unproven interventions. Various stakeholders, including governments, patient groups, medical societies, and the media, are committed to this outcome. In this perspective, we draw on our experience gained from partnerships in developing regenerative medicine products to identify technical, organizational, and ethical benchmarks for the responsible delivery of regenerative medicine treatments. These benchmarks may serve as the basis for policy interventions intended to drive the responsible delivery of stem cell and regenerative medicine products. Our particular focus is on a California‐based policy, but the suggested benchmarks are broadly applicable to national and international jurisdictions.

Almost 40 Years of Tissue Engineering in Russia: Where Are We Now?

This review describes achievements of Russian cell-based regenerative medicine in different periods of time depending on the legislation and politics, and future prospects for its commercialization and wide application with an emphasis on products devised for skin regeneration. The world’s experience in tissue engineering began with the development of living skin equivalents, utilizing a biopolymer matrix and cells at the very beginning of the 1980s. During this period, the USSR kept abreast with the times and also conducted studies on skin wound healing, implementing modern cell techniques. However, there soon emerged a gap between scientific advancement and practical application. After the breakup of the USSR, there were no institutions that could implement scientific inventions into full-scale manufacturing for clinical application. At the same time, accumulating scientific and practical experience allowed for the maintenance of biomedical research and its readiness for market entry at present. Recently developed legislation opens up new opportunities in this field in Russia. There are a growing number of studies on the development of novel products for regenerative medicine, bringing hope for its rapid progress.

Cryopreservation of Human iPS Cell Aggregates in a DMSO-Free Solution-An Optimization and Comparative Study.

Human induced pluripotent stem cells (hiPSCs) are an important cell source for regenerative medicine products. Effective methods of preservation are critical to their clinical and commercial applications. The use of a dimethyl sulfoxide (DMSO)-free solution containing all non-toxic molecules offers an effective alternative to the conventional DMSO and alleviates pain points associated with the use of DMSO in the cryopreservation of hiPSCs. Both hiPSCs and cells differentiated from them are commonly multicellular systems, which are more sensitive to stresses of freezing and thawing than single cells. In this investigation, low-temperature Raman spectroscopy visualized freezing behaviors of hiPSC aggregates in different solutions. These aggregates exhibited sensitivity to undercooling in DMSO-containing solutions. We demonstrated the ability to replace DMSO with non-toxic molecules, improve post-thaw cell survival, and reduce sensitivity to undercooling. An accelerated optimization process capitalized on the positive synergy among multiple DMSO-free molecules, which acted in concert to influence ice formation and protect cells during freezing and thawing. A differential evolution algorithm was used to optimize the multi-variable, DMSO-free preservation protocol in 8 experiments. hiPSC aggregates frozen in the optimized solution did not exhibit the same sensitivity to undercooling as those frozen in non-optimized solutions or DMSO, indicating superior adaptability of the optimized solution to different freezing modalities and unplanned deviations. This investigation shows the importance of optimization, explains the mechanisms and advantages of a DMSO-free solution, and enables not only improved cryopreservation of hiPSCs but potentially other cell types for translational regenerative medicine.

A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application.

IntroductionCultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine products and save many patients. For this purpose, establishing and disseminating the stale protocol of fabricating the cell sheet is crucial. The fabrication of cultured stratified epithelial cell sheets consists of many important steps, and since the patients’ epithelial cell conditions vary widely and are sometimes unstable, the qualities of the epithelial cell grafts are likewise potentially unstable. Therefore, in this paper, we report the stable protocol for fabrication of the transplantable cell sheet particularly from patient-derived oral mucosal tissues.MethodsSerum extracted from blood and buccal mucosal tissue were collected in Nagasaki University and transported to Tokyo Women's Medical University. Oral mucosal epithelial cells were collected by minimum trypsin method, and this treatment was studied whether to be a critical procedure. After 14 days cultivation, cultured cells were examined whether to be transplantable as cell sheets.ResultsWe successfully transported buccal mucosal tissue and serum without damage and contamination. Oral mucosal epithelial cells were collected with high viability by minimum trypsin method. Finally, we succeeded to stably fabricate oral mucosal epithelial cell sheets in all 10 patients.ConclusionsWe established a stable protocol for the fabrication of human oral mucosal epithelial cell sheets and their transportation in clinical settings in this study. These methodologies could also be basis for transplantation therapy using cultured cell sheets of various types other than oral mucosal epithelial cell and will contribute largely to the future development of regenerative medicine.

Medical Devices: Definition, Classification, and Regulatory Implications.

We propose the following definition of a medical device: "A contrivance designed and manufactured for use in healthcare, and not solely medicinal or nutritional." Current regulatory classifications of medical devices are complex and designed primarily for regulators. We propose a simpler classification, based on (1) the site of application of the device, (2) the time scale of its use, and (3) whether it has an external power source. The regulation of medical devices is less well developed than the regulation of medicinal products, which it could follow more closely. In particular, devices that incorporate medicines should be required to meet the same regulatory standards as medicinal products. This would remove the anomaly that some delivery systems that incorporate medicines are classified as devices while other similar systems that deliver the same medicines are classified as medicinal products. Some improvements might also result from more widespread use of registries, such as those used for prosthetic joint replacements. Registries would allow both a prospective examination of the performance of high-risk devices and a retrospective analysis when signals from other sources of information suggest problems. Those who apply for a marketing authorization for a new device should have to assure regulators of its quality of manufacture, safety, and efficacy before licensing. Even the most straightforward device should be shown to be useable in practice. Trials on patients, or at least simulations of use in the real world, should be practicable for most devices.

Therapeutic bacteriophages as a rescue treatment for drug-resistant infections - an in vivo studies overview.

Bacteriophages, highly prevalent in all environments, have found their use in medicine as an alternative or complement to antibiotics. The therapeutic use of bacteriophages was particularly popular in the 1920s and 1930s, until the discovery and introduction of antibiotics. Due to the dynamic growth of antibiotic resistance among bacterial strains, numerous international institutions (such as the FDA) have declared the search for novel treatment modalities to be of the highest priority. To date, bacteriophage therapy has not been registered for general use in Western countries. The regulation of biological medicinal products (within medicinal product regulation) does not contain a specific documentation frame for bacteriophages (only for vaccines, blood derived products, etc.) which, as active substances, need to meet specific requirements. Recently, the FDA allowed bacteriophage therapy to be used in the United States, via the Emergency Investigational New Drug scheme; clinical trials to compare the safety and efficacy of bacteriophage therapy are also permitted. To date, several therapeutic products of this type have made it to phase I or II; some clinical programmes have also been completed. This article cites numerous animal model studies and registered clinical trials, showing the safety and effectiveness of bacteriophage therapy, including infections caused by bacterial strains resistant to antibiotic treatment.

Plastic Products in Regenerative Medicine

Plastic Products in Regenerative Medicine

Effectiveness of autologous platelet-rich plasma gel in the treatment of hard-to-heal leg ulcers: a randomised control trial.

Regenerative medicine products such as autologous platelet-rich plasma (autologous PRP) gel may speed up the process of healing. Clinical studies show promising results in the treatment of diabetic foot ulcers (DFUs), however there is lack of scientific evidence of autologous PRP effectiveness in treating leg ulcers of other aetiology. This study evaluates the effectiveness of autologous PRP gel in the treatment of hard-to-heal leg ulcers compared with existing conventional treatment. A prospective, randomised controlled, open-labelled clinical trial was carried out between 2014 and 2018. An eight-week study protocol was chosen or until 100% wound re-epithelialisation was observed. Wound size reduction, granulation tissue formation, microbiological wound bed changes and safety were evaluated. A total of 69 patients (35 in the autologous PRP group and 34 in the control group) were included in the study; 25.71% of the autologous PRP group and 17.64% of control group had ulcers completely re-epithelialised (p>0.05). Wound size reduction in the autologous PRP group was 52.35% and 33.36% in the control group (p=0.003). The autologous PRP group showed superiority over conventional treatment in wound bed coverage with granulation (p=0.001). However, more frequent wound contamination was observed at the end of treatment in the autologous PRP group (p=0.024). No severe adverse events were noted during the study. Both treatment methods were considered equally safe. Topical application of autologous PRP gel in leg ulcers of various aetiology show beneficial results in wound size reduction and induces the granulation tissue formation. However, it is associated with more frequent microbiological wound contamination.

In vivo surveillance and elimination of teratoma-forming human embryonic stem cells with monoclonal antibody 2448 targeting annexin A2.

This study describes the use of a previously reported chimerised monoclonal antibody (mAb), ch2448, to kill human embryonic stem cells (hESCs) in vivo and prevent or delay the formation of teratomas. ch2448 was raised against hESCs and was previously shown to effectively kill ovarian and breast cancer cells in vitro and in vivo. The antigen target was subsequently found to be Annexin A2, an oncofetal antigen expressed on both embryonic cells and cancer cells. Against cancer cells, ch2448 binds and kills via antibody‐dependent cell‐mediated cytotoxicity (ADCC) and/or antibody‐drug conjugate (ADC) routes. Here, we investigate if the use of ch2448 can be extended to hESC. ch2448 was found to bind specifically to undifferentiated hESC but not differentiated progenitors. Similar to previous study using cancer cells, ch2448 kills hESC in vivo either indirectly by eliciting ADCC or directly as an ADC. The treatment with ch2448 post‐transplantation eliminated the in vivo circulating undifferentiated cells and prevented or delayed the formation of teratomas. This surveillance role of ch2448 adds an additional layer of safeguard to enhance the safety and efficacious use of pluripotent stem cell‐derived products in regenerative medicine. Thereby, translating the use of ch2448 in the treatment of cancers to a proof of concept study in hESC (or pluripotent stem cell [PSC]), we show that mAbs can also be used to eliminate teratoma forming cells in vivo during PSC‐derived cell therapies. We propose to use this strategy to complement existing methods to eliminate teratoma‐forming cells in vitro. Residual undifferentiated cells may escape in vitro removal methods and be introduced into patients together with the differentiated cells.

Examining Resources, Initiatives, and Regulatory Pathways to Advance Regenerative Medicine Manufacturing

The purpose of this review is to provide perspectives and resources for how cell and tissue manufacturing is being advanced by different stakeholders ranging from government to academia to even at a global scale. This perspective article illustrates resources and national efforts that are underway to advance cell and tissue manufacturing. Some of the efforts highlighted are only in their initial years so tangible results are still forthcoming. This perspective article is inspired by a workshop entitled “Cell and Tissue Manufacturing Innovation” that was held at the Tissue Engineering and Regenerative Medicine Society (TERMIS). Five main topics are discussed: (1) the US National Academies’ Forum on Regenerative Medicine, (2) academic translational programs, (3) development of measurements for regenerative medicine, (4) development of standards for ancillary materials, and (5) the global landscape for accelerating regenerative medicine manufacturing. Concepts are presented with current perspective and key insights on challenges, opportunities, and next steps for cell and tissue manufacturing innovation. Several insights are provided that may advance the manufacturing of regenerative medicine products in the USA.

The FDA's Regulatory Framework for Chimeric Antigen Receptor-TCell Therapies.

The FDA's Regulatory Framework for Chimeric Antigen Receptor-TCell Therapies.