Stem Cell Bank Research Articles

Induced Pluripotent Stem Cell-Derived Corneal Cells: Current Status and Application.

Deficiency and dysfunction of corneal cells leads to the blindness observed in corneal diseases such as limbal stem cell deficiency (LSCD) and bullous keratopathy. Regenerative cell therapies and engineered corneal tissue are promising treatments for these diseases [1]. However, these treatments are not yet clinically feasible due to inadequate cell sources. The discovery of induced pluripotent stem cells (iPSCs) by Shinya Yamanaka has provided a multitude of opportunities in research because iPSCs can be generated from somatic cells, thus providing an autologous and unlimited source for corneal cells. Compared to other stem cell sources such as mesenchymal and embryonic, iPSCs have advantages in differentiation potential and ethical concerns, respectively. Efforts have been made to use iPSCs to model corneal disorders and diseases, drug testing [2], and regenerative medicine[1]. Autologous treatments based on iPSCs can be exorbitantly expensive and time-consuming, but development of stem cell banks with human leukocyte antigen (HLA)- homozygous cell lines can provide cost- and time-efficient allogeneic alternatives. In this review, we discuss the early development of the cornea because protocols differentiating iPSCs toward corneal lineages rely heavily upon recapitulating this development. Differentiation of iPSCs toward corneal cell phenotypes have been analyzed with an emphasis on feeder-free, xeno-free, and well-defined protocols, which have clinical relevance. The application, challenges, and potential of iPSCs in corneal research are also discussed with a focus on hurdles that prevent clinical translation.

A study to assess the effectiveness of self instructional module to increase awareness of umbilical cord blood stem cell banking among eligible couples in selected urban community of Indore city in the year 2014-15

A study to assess the effectiveness of self instructional module to increase awareness of umbilical cord blood stem cell banking among eligible couples in selected urban community of Indore city in the year 2014-15

Generation of integration-free induced pluripotent stem cell line (KSCBi012-A) from urinary epithelial cells of a healthy male individual

A human induced pluripotent cell (hiPSC) line, KSCBi012-A, was generated from a 40-year-old male individual using non-integrating episomal vectors expressing reprogramming factors. The generated hiPSCs were integration-free, expressed pluripotency markers, exhibited the potential for differentiation into three germ layers in vivo, and maintained the normal karyotype. This cell line can be used as a control for a disease model and is available from Korea National Stem Cell Bank.

Biobanking of human pluripotent stem cells in China.

In recent years, significant progress has been made internationally in the development of human pluripotent stem cell (hPSC)‐derived products for serious and widespread disorders. Biobanking of the cellular starting materials is a crucial component in the delivery of safe and regulatory compliant cell therapies. In China, key players in these developments have been the recently launched National Stem Cell Resource Center (NSCRC) and its partner organizations in Guangzhou and Shanghai who together, have more than 600 hPSC lines formally recorded in the Chinese Ministry of Science and Technology's stem cell registry. In addition, 47 of these hPSCs have also been registered with the hPSCreg project which means they are independently certified for use in European Commission funded research projects. The NSCRC are currently using their own cell lines to manufacture eight different cell types qualified for clinical use, that are being used in nine clinical studies for different indications. The Institute of Zoology at the Chinese Academy of Sciences (IOZ‐CAS) has worked with NSCRC to establish Chinese and international standards in stem cell research. IOZ‐CAS was also a founding partner in the International Stem Cell Banking Initiative which brings together key stem cell banks to agree minimum standards for the provision of pluripotent stem cells for research and clinical use. Here, we describe recent developments in China in the establishment of hPSCs for use in the manufacture of cell therapies and the significant national and international coordination which has now been established to promote the translation of Chinese hPSC‐based products into clinical use according to national and international standards.

Construction strategies and prospect of the Global Medicinal Plant Stem Cell Bank

Medicinal plant stem cells are separated from the meristem and vascular cambium of medicinal plants, which can produce active components for preventing and treating diseases and improving body physical functions under certain conditions. Medicinal plant stem cells come from a broad category of medicinal plants, including ethnic medicinal plants, folk medicinal plants, original plants of health products, vegetables, fruits, and other potential medicinal plants. At present, the techniques for the isolation, identification, preservation and culture of medicinal plant stem cells have become increasingly mature, and the mechanism of stem cell differentiation, growth and regulation of secondary metabolites has been studied in depth. Medicinal plant stem cells have a broad application prospect in medicine, health food, food and medical beauty products. As a strategic resource, the construction of the "Global Medicinal Plant Stem Cell Bank" was first proposed to preserve various kinds of medicinal plant resources in the world, and it will go global relying on the internationalization strategy of traditional Chinese medicine. The bank should follow safety, environmental protection, advanced and practical design principles. The main construction contents include the original plant bank, stem cell bank, component resource bank, gene bank, database and resource sharing system, with genetic and data resources incorporated into the scope of protection and utilization. The bank will establish a new strategy for medicinal plant resources protection and regeneration, and provide a new resource for natural products drug discovery and a technology sharing platform for various medicinal plant stem cells. As a resource treasury, a source of innovative technologies and a center of cooperation, it will become the core driving force of the global medicinal plant stem cell industry.

Assessment of safety and immunogenicity of MHC homozygous iPSC-derived CD34+ hematopoietic progenitors in an NHP model

AbstractAdministration of ex vivo expanded somatic myeloid progenitors has been explored as a way to facilitate a more rapid myeloid recovery and improve overall survival after myeloablation. Recent advances in induced pluripotent stem cell (iPSC) technologies have created alternative platforms for supplying off-the-shelf immunologically compatible myeloid progenitors, including cellular products derived from major histocompatibility complex (MHC) homozygous superdonors, potentially increasing the availability of MHC-matching cells and maximizing the utility of stem cell banking. However, the teratogenic and tumorigenic potential of iPSC-derived progenitor cells and whether they will induce alloreactive antibodies upon transfer remain unclear. We evaluated the safety and efficacy of using CD34+CD45+ hematopoietic progenitors derived from MHC homozygous iPSCs (iHPs) to treat cytopenia after myeloablative hematopoietic stem cell (HSC) transplantation in a Mauritian cynomolgus macaque (MCM) nonhuman primate (NHP) model. We demonstrated that infusion of iHPs was well tolerated and safe, observing no teratomas or tumors in the MCMs up to 1 year after HSC transplantation and iHP infusion. Importantly, the iHPs also did not induce significant levels of alloantibodies in MHC-matched or -mismatched immunocompetent MCMs, even after increasing MHC expression on iHPs with interferon-γ. These results support the feasibility of iHP use in the setting of myeloablation and suggest that iHP products pose a low risk of inducing alloreactive antibodies.

Assessment of Knowledge and Attitude of Maternity Nurses regarding Umbilical Cord Stem Cells Collection and Banking

Assessment of Knowledge and Attitude of Maternity Nurses regarding Umbilical Cord Stem Cells Collection and Banking

Minimal Cube Explant Provides Optimal Isolation Condition of Mesenchymal Stem Cells from Umbilical Cord.

Enzymatic digestion and explant method have been widely used for isolating umbilical cord-derived mesenchymal stem cells (UC MSCs), although there is still a strong need for robust protocols for optimal isolation for large-scale stem cell banks. This study aims to establish an explant method for clinical scale production of MSCs from human UC tissue and to characterize UC MSCs isolated and cultured with the explant method. UC MSCs were isolated by enzymatic digestion, minimal cube explant (MCE) 1-2, MCE 2-4, and MCE 10 and cultured, respectively. Also, human antibody array and basic fibroblast growth factor (bFGF) secretion in conditioned medium (CM) was analyzed. The cells were evaluated initial cell number, colony forming unit-fibroblast (CFU-F), proliferation capacity, CD marker expression, and multi-lineage differentiation. SA-β-gal assay as well as expression of p16, p21 and p53 was performed by RT-PCR. MCE 2-4 is the most optimized method for isolation of small umbilical cord-derived fast proliferating cells (smumf cells) with the greatest number. MCE 2-4 had the highest secretion of various bioactive factors including bFGF. The MCE 2-4 provided significantly higher CD146 expression than enzymatic digestion, and that expression was maintained until P20. The gene expression of p16, p21, and p53 of smumf cells did not change until P10 and SA-β-gal activity did not increase until P14. This study demonstrated that MCE 2-4 provided an optimal environment to isolate MSCs with quantity and quality from human whole UC tissue through secretion of various bioactive factors inherent to UC.

Knowledge and Attitude of Taif University Students on Stem Cell Banking

Abstract: Background and Objectives: The awareness of university students is pivotal to the adaptation of new therapeutic technologies in combating illness and disorders. Stem cells are unique and effective in treating several pathological conditions. This study was an effort to determine the knowledge and attitude of Taif University students toward the banking of stem cells through the donation of umbilical cord blood (UCB). Materials and Methods: The study used a self-administered questionnaire that consisted of items on demographics, knowledge characteristics, and attitude towards banking of UCB as a source of stem cells for the purpose of achieving advanced therapeutic outcomes. The questionnaire was administered to only female students at Taif University during the academic year 2020-21. Both descriptive and inferential analysis were done using SPSSIBM 25 to analyze the data. Results: A total of 604 female students of Taif University responded to our questionnaire. Although most of the participants (70%) lacked sufficient knowledge (less than or equal to 2 out of 4), their attitudes were motivating, with more than 60% rated better than adequate (more than or equal to 3 out of 4). Many study participants have shown an interest in donating umbilical cord stem cells to treat other patients and for research purposes. Conclusion: Students with good knowledge of umbilical cord blood donation and those who are more educated showed a greater level of willingness to participate in banking and donation of stem cells. Therefore, raising awareness about stem cell banking and its importance among young undergraduates, especially female students, is imperative. Key words: Knowledge, Attitude, Taif University, Awareness campaigns, Umbilical Cord blood.

Knowledge, Attitude and Practice Regarding Dental Stem Cells and Tooth Banking among Dental Professionals: A Questionnaire-based Cross-sectional Study

Introduction: Stem cell therapy is new revolutionary technique which can repair many defects of human body. There are two sources of stem cells, one is embryonic and postnatal. Postnatal sites are the ones from which procuring stem cells is easiest. Dental stem cells from exfoliated deciduous teeth are also obtained from these sources. Dental health professionals must have understanding about dental stem cells for its application. Hence, there is need to evaluate knowledge, attitude and practice of dental professionals regarding dental stem cells. Aim: To evaluate knowledge, attitude and practice regarding dental stem cells and tooth banking among dental professionals of Gujarat. Materials and Methods: The study was a questionnaire based cross-sectional study conducted in the Department of Paediatric and Preventive Dentistry, KM Shah Dental College and Hospital, Sumandeep Vidyapeeth, Vadodara, Gujarat, India from December 2020 to March 2021. Total of dental health professionals participated including undergraduate (interns) and postgraduate students, dental practitioners and faculty members of dental colleges. Participants received the questionnaire in the form of a google form through email and whatsApp. The data was analysed using percentages of total and Chi-square test for statistical analysis. Results: Out of 711 participants, 410 were students, 44 faculties, 207 practitioners and 50 faculties and practitioner both. Among the faculty, knowledge on types of dental stem cells was significantly associated with the years of experience (p-value <0.001). Among the students, first year postgraduate students and interns had significantly less knowledge regarding various applications of dental stem cells (p-value <0.001). Among them, 52.9% participants were aware of stem cell banks in India and 55.5% were aware of names of stem cell banks. Participants suggested that seminars, journals, academic curriculum, conferences or Continued Dental Education (CDE), and advertisement and public notice all should be used to increase awareness of dental stem cells among dental professionals. Conclusion: There was adequate awareness among dental health professionals regarding dental stem cells and their applications but were unaware about tooth banking and its procedure. The study also found that there was lack of knowledge regarding steps of preservation of dental stem cells and their ethical concerns. Therefore, the topic of dental stem cells should be included in dental curriculum.

Knowledge and Attitude of Taif University Students on Stem Cell Banking

Abstract: Background and Objectives: The awareness of university students is pivotal to the adaptation of new therapeutic technologies in combating illness and disorders. Stem cells are unique and effective in treating several pathological conditions. This study was an effort to determine the knowledge and attitude of Taif University students toward the banking of stem cells through the donation of umbilical cord blood (UCB). Materials and Methods: The study used a self-administered questionnaire that consisted of items on demographics, knowledge characteristics, and attitude towards banking of UCB as a source of stem cells for the purpose of achieving advanced therapeutic outcomes. The questionnaire was administered to only female students at Taif University during the academic year 2020-21. Both descriptive and inferential analysis were done using SPSSIBM 25 to analyze the data. Results: A total of 604 female students of Taif University responded to our questionnaire. Although most of the participants (70%) lacked sufficient knowledge (less than or equal to 2 out of 4), their attitudes were motivating, with more than 60% rated better than adequate (more than or equal to 3 out of 4). Many study participants have shown an interest in donating umbilical cord stem cells to treat other patients and for research purposes. Conclusion: Students with good knowledge of umbilical cord blood donation and those who are more educated showed a greater level of willingness to participate in banking and donation of stem cells. Therefore, raising awareness about stem cell banking and its importance among young undergraduates, especially female students, is imperative. Key words: Knowledge, Attitude, Taif University, Awareness campaigns, Umbilical Cord blood.

Effects of gestational diabetes mellitus on the quality and quantity of blood hematopoietic stem cells: a case-control study

AimTo evaluate the effects of gestational diabetes mellitus (GDM) on the quantity and quality of hematopoietic stem cells (HSC).MethodsIn this case-control study, HSC were isolated from umbilical cord blood (UCB) procured at delivery from 63 mothers with GDM and 67 healthy mothers. Total nucleated cells (TNC) and CD34+ cells were quantified using BD FACSCalibur flow cytometer. The quantity and quality of stem cells were determined.ResultsThe GDM group had lower total cord blood volume and lower number of nucleated HSC compared with healthy mothers. Regarding stem cell quantity parameters, they had significantly lower UCB volume (P = 0.041), TNC count (P = 0.022), total viable NC count (P = 0.014), and CD34+ percentage (P = 0.014). Regarding the quality of stem cells, they had significantly lower viable TNC percentage (P = 0.015). The predictors for total TNC count were longer labor duration (adjusted B coefficient [p]: 0.031 [0.046]), greater estimated blood loss (0.089 [0.005]), female neonates (12.322 [0.049]), and higher placenta weight (0.080 [0.033]). The predictors of total viable NC count were greater estimated blood loss (0.092 [0.003]), female neonates (13.16 [0.035]), and greater placenta weight (0.083 [0.026]).ConclusionThe GDM group had much lower quantity and quality of UCB stem cells. Our results should be taken into consideration when drawing cord blood for unrelated stem cell banking in an obstetric unit to ensure the obtaining of optimal cord blood samples and to avoid unnecessary expenses.

A study to assess the level of knowledge regarding umbilical cord stem cell banking among staff nurses at selected Hospitals of Haridwar Uttrakhand

A descriptive study was conducted to assess the level of knowledge regarding umbilical cord stem cell banking among staff nurses at selected Hospitals of Haridwar Uttrakhand A non-experimental research design was used assess the knowledge regarding umbilical cord stem cell banking among staff nurses. The sample consists of 104 staff nurses of selected hospitals who were available at the time of study. Purposive sampling technique was used for selection of samples. The data was collected using self-structured questionnaire. Results depicted that majority of staff nurses had inadequate knowledge regarding umbilical cord stem cell banking. The mean knowledge score was found to be 16.5±6.16. The results also indicated knowledge of staff nurses was found to be significant with years of experience of staff nurses. Hence the study concluded that knowledge of staff nurses were influenced by years’ experience

Effect of Instructional Guidelines on Antenatal Mothers' Knowledge regarding Umbilical Cord Stem Cell Banking

Effect of Instructional Guidelines on Antenatal Mothers' Knowledge regarding Umbilical Cord Stem Cell Banking

A STUDY TO ASSESS THE EFFECTIVENESS OF PLANNED TEACHING PROGRAMME REGARDING STEM CELLS AMONG ELIGIBLE COUPLES IN SELECTED AREAS OF AHMEDABAD

Introduction: Stem cells have tremendous promise to helping us to understand and treat a range of various diseases, injuries and other health related conditions. Their potential is evident in the use of cord blood stem cells to treat diseases of the blood, A cord blood stem cells therapy has saved the lives of thousands of children with leukemia and can be seen in the use of stem cells for tissue grafts to treat diseases or injury to the bone, skin and surface of the eye. Materials and Methods: This pre-experimental study was conducted on eligible couples with sample size of age between 15-45 years, willingness to participate in study in selected areas of Ahmedabad. The investigator adopted purposive technique to select sample. Data was collected using 4 demographic variables & 20 questionnaires regarding stem cells, its uses, diseases treated by stem cells & stem cell banking were included. A pre-test was conducted on 40 samples after which the planned teaching programme was implemented followed by the post test. Result: Data gathered was analyzed and interpreted using both experimental and inferential statistics. The mean and SD of pre-test was 5.725 and 1.3957, whereas the mean and SD of post- test was 16.25 and 1.1036, the calculated t value was greater than tabulated t value. Hence the null hypothesis was rejected and the research hypothesis was accepted. The result shows that when planned teaching program was given to eligible couples age group people, they achieve the best scores. Conclusion: This Study intends to assess the effectiveness on plan teaching program regarding stem cells among eligible couple in selected areas of Ahmedabad. This Study reveals that the post-test knowledge score is higher than the pre-test knowledge score regarding stem cells storage, stem cells banking among the eligible couple.

Infusion of iPSC-Derived CD34 + Hematopoietic Progenitors Following Myeloablative HSC Transplantation and Assessment of Their Immunogenicity in MHC-Defined Nonhuman Primate Model

Bone marrow suppression and associated neutropenia that renders the patient more susceptible to infection is a major complication and limiting factor for current chemotherapy. Strategies aimed at neutrophil reconstitution using transfusions or accelerating neutrophil recovery with G-CSF show a limited effect in reducing the rate of infection. To overcome these limitations, administration of ex vivo expanded somatic hematopoietic progenitors (HPs) has been explored as a way to facilitate a more rapid myeloid recovery and improve overall survival following myeloablation. Recent advances in reprogramming and induced pluripotent stem cell (iPSC) technologies have created alternative platforms for off-the-shelf supply of immunologically compatible HPs, including cellular products derived from MHC homozygous superdonors which can increase the degree of MHC matching and may provide a maximum utility for stem cell banking. In these studies, we developed Mauritian cynomolgus macaque (MCM) model to evaluate the utility and safety of CD34 +CD45 + hematopoietic progenitors derived from iPSCs (iHPs) generated from MHC homozygous animals in the treatment of cytopenia following myeloablative stem cell transplantation. MCM iPSCs were generated from MHC homozygous animals and used to generate iHPs in coculture with OP9. Three groups of MCMs underwent myeloablative total body irradiation (TBI) followed by transplantation of cryopreserved autologous CD34 + HSPCs. Four days after autologous HSPC transplantation, animals received transfusion of 30x10 6/kg cryopreserved iCD34 + cells tagged with eGFP or tdTomato from homozygous MHC-matched iPSCs (group 2) or homozygous MHC mismatched iPSCs (group 3). We have demonstrated that infusion of iHPs is safe and well tolerated. No teratoma or tumor formation was observed in animals one year after infusion of iHPs. Although we detected few iPSC derived hematopoietic cells in the blood within 1-week post-infusion, we did not see any significant differences in blood counts or other peripheral blood parameters between all animal groups. Intriguingly we found footprints of iPSC-derived cells in the colon, lymph node, skin and brain specimens collected 25 days after iHP infusion from one animal. This suggests a possibility of migration and retention within these tissues of iHP and iHP-derived myeloid cells. To assess immunogenicity, MHC homozygous iHPs were infused into immunocompetent MCMs across different MHC barriers. 10x10 6/kg of cryopreserved iCD34+ from homozygous iPSCs were transfused every week for 3 weeks, into animals that were divided into 4 groups namely autologous (group I), MHC matched homozygous (group II), MHC matched heterozygous (group III) and MHC mismatch (group IV). Overall, these studies revealed low immunogenicity of MHC homozygous iHPs in MHC matched homozygous and heterozygous animals, while weak immune response was detected following iHP infusion in some MHC mismatched animals. DisclosuresSlukvin: Cynata Therapeutics: Consultancy, Current equity holder in publicly-traded company.

Cryoprotective Effect of Pentoxifylline on Spermatogonial Stem Cell During Transplantation into Azoospermic Torsion Mouse Model.

Preserving the spermatogonial stem cells (SSCs) in long periods of time during the treatment of male infertility using stem cell banking systems and transplantation is an important issue. Therefore, this study was conducted to develop an optimal cryopreservation protocol for SSCs using 10mM pentoxifylline (PTX) as an antioxidant in basal freezing medium. Testicular torsion-a mouse model for long-term infertility-was used to transplant fresh SSCs (n = 6), fresh SSCs treated with PTX (n = 6), cryopreserved SSCs with basal freezing medium (n = 6), and cryopreserved SSCs treated with PTX (n = 6). Eight weeks after germ cell transplantation, samples were assessed for proliferation, through evaluation of Ddx4 and Id4 markers, and differentiation via evaluation of C-Kit and Sycp3, Tnp1, Tnp2, and Prm1 markers. According to morphological and flow cytometry results, SSCs are able to form colonies and express Gfra1, Id4, α6-integrin, and β1-integrin markers. We found positive influence from PTX on proliferative and differentiative markers in SSCs transplanted to azoospermic mice. In the recipient testis, donor SSCs formed spermatogenic colonies and sperm. Respecting these data, adding pentoxifylline is a practical way to precisely cryopreserve germ cells enriched for SSCs in cryopreservation, and this procedure could become an efficient method to restore fertility in a clinical setup. However, more studies are needed to ensure its safety in the long term.

Marketing and Survey Analysis of Stem Cell Banking in Western Maharashtra

Purpose to check the knowledge of general population and doctors from western Maharashtra about the basics, purpose, utility and capability of stem cells banking and future scopes in medical field. Data was collected from three hospitals from Sangali district. For given study one thousand and two hundred individual from different background and different age group participated between 28th October to 4th January. Among these people 64% of general population heard about stem cell and cord blood banking, only 20% people have detailed information of stem cell banking and 70% medical staff have positive approach toward stem cell banking. Majority of people don’t know importance of cells on deadly disease. Lot of people unaware about technical terms and preservation techniques. Doctors have different opinion on use of stem cells on genetic disorder. 30% doctors though still it need more research and development for use of medical purpose. Hospitals, Institute and young generation need to take action on awareness of stem cell banking. obstetricians and the pediatricians should take a central role in providing the correct information to would be parents to help them in taking a correct decision.

Contributors

Robin Gillis Emeritus Professor of Applied Theology at the University of Kent and an Honorary Provincial Canon of Canterbury Cathedral and Canon Theologian (and for three years Acting Dean) of Holy Trinity Cathedral, Gibraltar. For thirteen years he was chair of the Archbishop of Canterbury’s Medical Ethics Advisory Group. He is a member of the British Medical Association’s Medical Ethics Committee, of the Medical Research Council’s Stem Cell Bank Steering Committee and, previously, of the Nuffield Council on Bioethics. He is editor of the journal Theology and author or editor of some forty books on Christian ethics

Evaluation of the Spanish population coverage of a prospective HLA haplobank of induced pluripotent stem cells

BackgroundiPSC (induced pluripotent stem cells) banks of iPSC lines with homozygous HLA (human leukocyte antigen) haplotypes (haplobanks) are proposed as an affordable and off-the-shelf approach to allogeneic transplantation of iPSC derived cell therapies. Cord blood banks offer an extensive source of HLA-typed cells suitable for reprogramming to iPSC. Several initiatives worldwide have been undertaken to create national and international iPSC haplobanks that match a significant part of a population.MethodsTo create an iPSC haplobank that serves the Spanish population (IPS-PANIA), we have searched the Spanish Bone Marrow Donor Registry (REDMO) to identify the most frequently estimated haplotypes. From the top ten donors identified, we estimated the population coverage using the criteria of zero mismatches in HLA-A, HLA-B, and HLA-DRB1 with different stringencies: high resolution, low resolution, and beneficial mismatch.ResultsWe have calculated that ten cord blood units from homozygous donors stored at the Spanish cord blood banks can provide HLA-A, HLA-B, and HLA-DRB1 matching for 28.23% of the population.ConclusionWe confirm the feasibility of using banked cord blood units to create an iPSC haplobank that will cover a significant percentage of the Spanish and international population for future advanced therapy replacement strategies.