Stem Cell Bank Research Articles

Comparison of umbilical cord tissue-derived mesenchymal stromal cells isolated from cryopreserved material and extracted by explantation and digestion methods utilizing a split manufacturing model

Background aimsUmbilical cord (UC) tissue is recognized as an advantageous source of mesenchymal stromal cells (MSCs), whose therapeutic properties are being actively evaluated in pre-clinical and clinical trials. In recognition of its potential value, storage of UC tissue or cells from UC tissue in newborn stem cell banks is now commonplace; however, strategies for isolating UC-derived MSCs (UCMSCs) from UC tissue have not been standardized. The majority of newborn stem cell banks take one of two approaches to cord tissue processing and cryopreservation: enzymatic digestion of the fresh tissue with cryopreservation of the subsequent cell suspension or cryopreservation of the tissue as a composite whole with later, post-thaw isolation of cells by explantation. Evaluation of UCMSCs derived by these two principal preparation and cryopreservation strategies is important to understanding whether the methods currently employed by newborn stem cell banks retain the desirable clinical attributes of UC cells. MethodsUCMSCs were isolated from 10 UC tissue samples by both explantation and enzymatic digestion methods to allow for comparison of cells from the same donor. Cell isolates from both methods were compared pre- and post-cryopreservation as well as after serial passaging. Cell viability, morphology, growth kinetics, immunophenotype, cytokine secretion and differentiation capacity were evaluated. ResultsUCMSCs could be derived from fresh UC tissue by both explantation and digestion methods and from thawed UC tissue by explantation. Initial cell populations isolated by digestion were heterogeneous and took longer to enrich for UCMSCs in culture than populations obtained by explantation. However, once isolated and enriched, UCMSCs obtained by either method showed no significant difference in viability, morphology, rate of proliferation, surface marker expression, levels of cytokine secretion or differentiation capacity. ConclusionsDerivation of UCMSCs by explantation after thawing UC cryopreserved as a composite tissue may be favorable in terms of initial purity and number of cells achievable by a specific passage. However, we observed no evidence of functional difference between UCMSCs derived by explanation or digestion, suggesting that cells isolated from cryopreserved material obtained by either method maintain their therapeutic properties.

Stem Cell Statute in Jordan: Leading the Way

The use of stem cells in research has caused much controversy and ethical dilemma. The primary source of stem cells is human embryos, a source which has been confronted with objections based on ethical, moral, and religious positions. Jordan has passed the first of its-kind Statute in the region, aiming at regulating the use of stem cells both for therapeutic and research purposes. The Statute adopted a regulatory approach between the restrictive and intermediate. The Statute, however, pays more attention to stem cell banking in many of its articles. Many critical aspects in regulating stem cell research activities are overlooked. This is including but not limited to the process of informed consent, protecting privacy, maintaining confidentiality, the need for a national entity responsible for regulating embryonic stem cell (ESC) research, and requirements of monitoring activity. The authors recommend further review of the current Statute in light of the deficiencies discussed so as to develop a more comprehensive and coherent Statute.

Type I collagen facilitates safe and reliable expansion of human dental pulp stem cells in xenogeneic serum-free culture

BackgroundHuman dental pulp stem cells (DPSCs) are a readily accessible and promising cell source for regenerative medicine. We recently reported that a xenogeneic serum-free culture medium (XFM) is preferable to fetal bovine serum-containing culture medium for ex vivo expansion of DPSCs; however, we observed that, upon reaching overconfluence, XFM cells developed a multilayered structure and frequently underwent apoptotic death, resulting in reduced cell yield. Therefore, we focused on optimization of the XFM culture system to avoid the undesirable death of DPSCs.MethodsWe selected type I collagen (COL) as the optimal coating substrate for the cultureware and compared DPSCs cultured on COL in XFM (COL-XFM cells) to the conventional XFM cultures (XFM cells).ResultsOur results demonstrated that COL coating facilitated significantly higher rates of cell isolation and growth; upon reaching overconfluence, cell survival and sustained proliferative potential resulted in two-fold yield compared to the XFM cells. Surprisingly, after subculturing the overconfluent COL-XFM cultures, the cells retained stem cell behavior including stable cell growth, multidifferentiation potential, stem cell phenotype, and chromosomal stability, which was achieved through HIF-1α-dependent production and uniform distribution of collagen type I and its interactions with integrins α2β1 and α11β1 at overconfluency. In contrast, cells undergoing apoptotic death within overconfluent XFM cultures had disorganized mitochondria with membrane depolarization.ConclusionThe use of COL as a coating substrate promises safe and reliable handling of DPSCs in XFM culture, allowing translational stem cell medicine to achieve stable isolation, expansion, and banking of donor-derived stem cells.

Assess the effectiveness of video assisted teaching programme on umbilical cord blood stem cell collection and banking among BSc (N) III year students in SMCH

Assess the effectiveness of video assisted teaching programme on umbilical cord blood stem cell collection and banking among BSc (N) III year students in SMCH

How old is too old? In vivo engraftment of human peripheral blood stem cells cryopreserved for up to 18 years - implications for clinical transplantation and stability programs.

Peripheral blood stem cells (PBSC) are commonly cryopreserved awaiting clinical use for hematopoietic stem cell transplant. Long term cryopreservation is commonly defined as five years or longer, and limited data exists regarding how long PBSC can be cryopreserved and retain the ability to successfully engraft. Clinical programs, stem cell banks, and regulatory and accrediting agencies interested in product stability would benefit from such data. Thus, we assessed recovery and colony forming ability of PBSC following long-term cryopreservation as well as their ability to engraft in NOD/SCID/IL-2Rγnull (NSG) mice. To investigate the in vivo engraftment potential of long-term cryopreserved PBSC units. PBSC units which were collected and frozen using validated clinical protocols were obtained for research use from the Cellular Therapy Laboratory at Indiana University Health. These units were thawed in the Cellular Therapy Laboratory using clinical standards of practice, and the pre-freeze and post-thaw characteristics of the units were compared. Progenitor function was assessed using standard colony-forming assays. CD34-selected cells were transplanted into immunodeficient mice to assess stem cell function. Ten PBSC units with mean of 17 years in cryopreservation (range 13.6-18.3 years) demonstrated a mean total cell recovery of 88% ± 12% (range 68%-110%) and post-thaw viability of 69% ± 17% (range 34%-86%). BFU-E growth was shown in 9 of 10 units and CFU-GM growth in 7 of 10 units post-thaw. Immunodeficient mice were transplanted with CD34-selected cells from four randomly chosen PBSC units. All mice demonstrated long-term engraftment at 12 wk with mean 34% ± 24% human CD45+ cells, and differentiation with presence of human CD19+, CD3+ and CD33+ cells. Harvested bone marrow from all mice demonstrated growth of erythroid and myeloid colonies. We demonstrated engraftment of clinically-collected and thawed PBSC following cryopreservation up to 18 years in NSG mice, signifying likely successful clinical transplantation of PBSC following long-term cryopreservation.

Comparison of functional attributes of mscs isolated by explant and digestion umbilical cord tissue using a split manufacturing model

Background & Aim Umbilical cord tissue (CT) is a rich source of mesenchymal stromal cells (MSCs) and many newborn stem cell banks have established programs for cryopreservation of CT as a source material for future clinical use. Here we utilize a split manufacturing model to compare MSCs isolated after thawing cord tissue cryopreserved as composite cord tissue (c-CT MSCs) and MSCs isolated from previously-cryopreserved, heterogeneous, single-cell suspensions of enzymatically-digested cord tissue (d-CT MSCs) to determine if growth kinetics and differentiation capacity are impacted by the two predominant CT storage strategies adopted by the banking industry. Methods, Results & Conclusion Methods: 10 umbilical cords were obtained for research from consenting mothers. Each CT was divided in half with one half cut into small pieces cryopreserved as whole, composite material, and the other half enzymatically digested to obtain a heterogeneous cell suspension that was cryopreserved. After at least 1 week, thawed composite tissue was explanted in MSC-selective medium for 2 weeks while cells from digested tissue were seeded directly into culture. Cells from both methods were expanded to the end of the third passage (P3) and assessed by flow cytometry, viability, and differentiation capacity. Results Cell yield/gr41 and post-thaw recovery were greater for explanted material (Table 1). Initial heterogeneity of d-CT is addressed by serial culture; P3 d-CT populations are enriched for MSCs, with no differences between CD73, CD90, and CD34/45 compared to c-CT populations (Figure 1). Viabilities and doubling times at P3 were similar (Table 2). Similar qualitative adipo-, and chondrogenic differentiation capabilities were observed for c-CT and d-CT cells while osteogenic capacity was confirmed with calcium deposition as well as an increase in ALP activity (2.7 and 2.9 fold increase for c-CT and d-CT, p=0.8). Conclusion The processes for cryopreservation and isolation of c-CT and d-CT MSCs differ, but once a purified population is obtained, MSCs from the same donor are indistinguishable with regards to growth kinetics and tri-lineage differentiation potential. These results build on our previous report that cells from the same donor are not notably different in secreted factors. While these studies highlight that either CT cryopreservation strategy is suitable for maintaining MSC properties, storage of composite tissue is advantageous due to the greater purity of cells and higher per-gram yield.

Karolinska Institutet Human Embryonic Stem Cell Bank.

The Karolinska Institutet Human Embryonic Stem Cell Bank (KI Stem Cell Bank) was established at KI, Stockholm, Sweden, when the first human embryonic stem cell (hESC) line was derived by Professor Hovatta and colleagues in 2002. Since then, the bank has grown to include 60 hESC lines. From the very beginning the aim of the bank has been derivation of hESC lines suitable for clinical use. Step by step progress has been made towards this goal, including removal of xeno components, establishment of chemically defined conditions and Good Manufacturing Practice (GMP) compliancy. Today our bank includes such clinical grade hESC line, KARO1, derived and banked according to GMP guidelines. Many of the hESC lines in the bank have been distributed to the scientific community and are deposited in the Stockholm Medical Biobank available for research on collaborative basis.

GMP Banking of Human Pluripotent Stem Cells: A US and UK perspective.

GMP Banking of Human Pluripotent Stem Cells: A US and UK perspective.

Governamentalidade neoliberal, estratégias e seus efeitos: problematizações em torno dos biobancos de células-tronco

O artigo visa abordar uma análise, em formato de ensaio analítico a respeito das práticas de guarda de células-tronco. A chamada medicina personalizada e regenerativa emerge ligada ao capitalismo neoliberal e a uma modalidade de governo das condutas baseada na biomedicina e em uma política de gestão da vida focada no mercado neoliberal securitário. A saúde foi transformada em um comércio de células e sustentada nas táticas da bioeconomia, do biovalor e das biotecnologias, articuladas em um dispositivo de governamentalidade reguladora do ser saudável, a partir da compra e venda de materiais biomoleculares e genéticos. Concluindo, busca-se interrogar o modo de organizar e funcionar dos biobancos públicos e privados de células-tronco do cordão umbilical e os efeitos destes mecanismos biopolíticos.

Knowledge, attitudes, and practices of undergraduate students concerning Regenerative Endodontics.

Knowledge, attitudes, and beliefs of the students are important for the Tissue Engineering in Endodontic practice. The opinion of these future dentists would ultimately will decide the endurance of REPs as routine procedures in endodontic practice. The aim of this study was to perform a survey to identify the knowledge, attitudes, and practices of undergraduate students about regenerative endodontic procedures (REPs). The questionnaire was obtained after cross-cultural adaptation of a questionnaire previously applied in USA and was applied to two hundred forty-eight undergraduates. Data were statistically analyzed. Most of the students (82.9%) agreed that regenerative therapy should be incorporated to dentistry and 87.5% of them believed that stem cells banks would be useful for the tissue regeneration. Most participants (58.1%) would like to obtain an internship/tutoring that addresses REPs and 80.8% of participants think that the major obstacle to a patient accepting a REP was the expected high cost of the treatment. The freshmen students were more optimistic about offering stem cell treatments to their patients (P≤0.05). The undergraduates were very optimistic about the future of REPs, stem cell banking, and tissue engineering. Although seniors demonstrated less enthusiasm towards REPs than the freshman, most students are willing to recommend these treatments to their patients.

Road to clinical implementation of CAR-T technology in Poznań

AbstractThe objective of this paper is to present the process of the national and international accreditation leading to the establishment of the first certified chimeric antigen receptor T (CAR-T) Cell Unit in Poland on the basis of the Department of Hematology and Bone Marrow Transplantation in Poznan University of Medical Sciences and first successful CAR-T therapy in Poland. During 12 months from the initial decision to establish the CAR-T Cell Unit to the application of CAR-T cell treatment in the first patient, the center had to undergo the multidisciplinary external and internal training, as well as the adaptation of multiple procedures within the Transplant Unit and Stem Cell Bank. In order to get accreditation for the implementation of CAR-T cell therapy, an initial training of the team involved in handling cellular products and patient care was organized and updated as a continuous process. The Department fulfilled the site-selection international criteria. The first patient diagnosed for refractory/relapsed DLBCL was qualified, and finally CAR-T cells were administered with successful clinical outcome.

Need for Specialized Therapeutic Stem Cells Banks Equipped with Tumor Regression Enzymes and Anti-Tumor Genes.

Stem cells are currently being used in many clinical trials for regenerative purposes. These are promising results for stem cells in the treatment of several diseases, including cancer. Nevertheless, there are still many variables which should be addressed before the application of stem cells for cancer treatment. One approach should be to establish well-characterized therapeutic stem cell banks to minimize the variation in results from different clinical trials and facilitate their effective use in basic and translational research.

Післязабійний кістковий мозок великої рогатої худоби – перспективне джерело стовбурових клітин

The development of modern methods of cryopreservation of animal cells has promoted to introduce the concept of creating of stem cell banks for their future use, to avoid problems with donor shortages. Large-scale cryopreservation of stem cells began in the 1990s with the establishment of human cord blood banks. The issue of long-term storage of stem cells is also acute in veterinary medicine. To obtain of cellular material, is needed a source suitable for cell isolation. As such a source it is advisable to consider a slaughter material from productive animals. The purpose of this study is to establish the possibility of cattle bone marrow use after slaughter as source of stem cells, on the basis of the proliferation index and the viability of the cultured cells to determine the suitability of this biological material for the obtaining of stem cells 72 hours after slaughter of the animal. Materials and methods. The bone marrow was obtained from the femur of a cow, 3 years old, who was slaughtered in a slaughterhouse. The bone marrow was collected with sterile tweezer into a sterile tube, filled with 0.25% trypsin solution (ratio of bone marrow volume to trypsin solution – 10: 1) and placed in a refrigerator (t + 4 0C) for 24 hours for enzymatic disaggregation. After bone marrow disaggregation, cultivation of cells was performed in a CO2-incubator using of disposable plastic Petri dishes (d = 30 mm) according to the standard procedure by periodically passaging them after forming a monolayer by 95–100%. Results. Cultivation of a suspension of cells, derived from the post-mortemal bone marrow of a cow, revealed that colonies of cells began to emerge 6–7 days after sowing. Passaging the cells with 0,25% trypsin-Versen solution and sowing them in new cultural plates contributed to increase the mass of cells that actively proliferated. It was established that stem cells, isolated from the post-mortemal bone marrow of cattle, have significant proliferative potential, as evidenced by proliferation indices  from I to III passages and high viability of cells. Thus, the post-mortemal bone marrow of the cattle can be used as an alternative source of stem cells. This biological material is suitable for the isolation of stem cells, even 72 hours after slaughtering the animal, which opens the prospects of its transportation over long distances.

A Study to Assess the Effectiveness of Structured Teaching Programme on Knowledge regarding Umbilical Cord Stem Cell Banking among Antenatal Mothers in selected Hospital, at Rajkot

A Study to Assess the Effectiveness of Structured Teaching Programme on Knowledge regarding Umbilical Cord Stem Cell Banking among Antenatal Mothers in selected Hospital, at Rajkot

Stem cells in the oral cavity – An overview

Stem cells play a vital role in regenerative medicine and it has the capacity to form any type of tissue under favored circumstances. The stem cells from the oral cavity can be easily removed and used for the regeneration of both hard and soft tissues. This article aimed to review the sources of stem cells from the oral cavity and the application of the same in the orofacial region and the principles of stem cell bank for the betterment of the quality and quantity of stem cells.

A prospective Saudi dental stem-cell bank from the perspective of the public and dental practitioners: A cross sectional survey.

Objectives:The aim of this study (1) To evaluate the Saudi public's knowledge and attitude on dental stem cells (DSC) for therapeutic/banking purposes, (2) To evaluate the perception of Saudi dentists towards DSC and their attitude towards banking of DSC.Methodology:This was a cross-sectional study based on an electronic survey distributed through the social media nationwide among the Saudi public, and through paper-based surveys among dentists in Riyadh. By convenience, eligible study participants were Saudi adults from the community and dentists from 17 dental centers in different regions of Riyadh. Using SPSS v. 25, descriptive statistics (n,%; PMS ± SD) and bivariate analyses (Pearson's Chi square, Mann Whitney) were conducted to determine factors associated with the study outcomes, with a P value statistically significant at <0.05.Results:For the Saudi public, 1494 participants completed the survey. The PMS ± SD of public knowledge was 25.5 ± 25.9, while their attitude was 80.2 ± 27.0. Factors associated with higher knowledge scores were younger age groups, while female participants, older age groups, university educated and employed participants had higher attitude. For Saudi dentists, 246/262 (94%) dentists responded to the questionnaire. Their PMS ± SD of perception towards DSC research for regenerative purposes was 74.5 ± 15.6. Factors associated with higher perception scores were those with more experience. Dentists who had higher perception scores towards DSCs were significantly more willing to save teeth for regenerative dental treatment.Conclusion:The Saudi public community had poor knowledge about the therapeutic and research benefits of DSC, yet high degree of attitude to enroll in a future Saudi DSC bank. Saudi dentists had moderately high levels of perception towards DSC research.

Assessment of Post-thaw Quality of Dental Mesenchymal Stromal Cells After Long-Term Cryopreservation by Uncontrolled Freezing.

Cryopreservation abilities of dental tissue-derived mesenchymal stromal cells (DMSCs) including dental pulp stem cells (DPSCs) and dental follicle stem cells (DFSC) play an important role in the applications of these cells in clinical settings. In this context, we checked whether storage at - 80°C in 10% DMSO for a longer period has any adverse effect on the functionality and genetic stability. We carried our studies on DPSC and DFSC samples that were revived after a maximum of 5years of cryopreservation. We observed that even after long-term uncontrolled freezing at - 80°C, these cells survived and proliferated efficiently. The assessment was made based on their post-thaw morphology, immunophenotypes, differentiation potential, growth kinetics, and genetic features. These cells retained the expression of stemness markers, differentiation ability and maintained their normal karyotype. Our results indicated no significant morphological or immunophenotypic differences between the cryopreserved DMSCs and the fresh DMSCs. Our study implies that mesenchymal stromal cells derived from the dental tissue origin are very robust and do not require any sophisticated preservation protocols. Thus, these can be an ideal source for research, stem cell banking, as well as successful clinical applications in tissue engineering and cell-based therapeutics. Graphical Abstract Schematic diagram showing the cryopreservation of DMSCs by uncontrolled freezing at -80 c has no adverse effects on their functionality and genetic stability.

"Dental Stem Cells": Awareness, knowledge, and attitude of dental professionals-A cross-sectional study.

The present study was conducted with an aim to assess the awareness about existence of dental stem cells among dental professionals. A total sample size of 100 dental professionals in the age group of 22-32years, comprising of 28 interns, 46 postgraduate students, and 26 senior residents from various specialties was selected. A self-administered, hand-delivered, 10-point questionnaire was used to assess the awareness, knowledge, and attitude of dental professionals towards dental stem cells. Results were documented and statistically analyzed using chi-square test. A total of 72% dental professionals were aware about dental stem cells with dental curriculum books being the main source of information. Note that 81% of positive respondents were aware about the different types and a total of 28% were aware about the procedure to procure dental stem cells. Only 33.3% of positive respondents were aware about existence of dental stem cell banks in India. A high level of awareness about dental stem cells and its applications was noted during the course of this study. A significant association of age and type of practice with awareness of dental stem cells and knowledge regarding their use in developing nondental tissues was observed. A positive attitude towards recommending dental stem cell banking was seen.

Factors affecting hematopoietic stem cells derived from umbilical cord blood

Background: Stem cells from umbilical cord blood Lifetime protection umbilical cord blood contains stem cells. These stem cells were until recently received and discarded after delivery with umbilical cord and placenta. However, these cells are of great value at the moment of birth and kept for life for any use. May be necessitated by urgent medical imperatives given later. Human stem cells derived from the umbilical cord have many advantages: they are easily collected without harm to the child or mother and treatment of many diseases including blood diseases, cancer and immune system disorders.Because of the high cost of treatment and preservation of umbilical cord blood and the relative proportion of stem cells in which the banks of stem cells from umbilical cord blood paid great attention to the acquisition of high-quality units to ensure higher rates of success of stem cell transplantation and so I study the factors that may affect the quality of umbilical cord blood units is important. Aim: To determine the characteristics of the laboratory hematopoietic potentiality of umbilical cord blood hematopoietic stem cells, and to study their association with maternal and neonatal factors. Materials & Methods: The maternal and neonatal factors that influence the total nucleated cell (TNC), and CFU yields in CB collected for the Cord Blood Bank were evaluated.Results: As expected, there was a significant rise in TNC / CBU and CFU / CBU considering quality of UCB unit with increase in UCB volume, and also there was significant elevation of cord blood volume, TNC x 108 / CBU and CFU / CBU x105 in units collected from birth weight ˃ 3500 gm compared to that of birth weight ≤ 3500 gm. Fetal gender affected quality of UCB units, Volume of UCB units in our study was significantly higher in male fetus compared to only in female fetus. TNC x 108 / CBU was elevated in preterm than collected from post term. CFU / CBU x 105 in preterm compared to term and post term. The increase in maternal age was associated with high volume, TNC and CFU. There is significant elevation of CBU volume, TNC , from maternal age > 20 years compared to that of maternal age ≤ 20 years. Conclusions: Our study suggests that maternal donor characteristics significantly influence the yield of TNC and viability of UCB samples. These factors should be considered when attempting to improve the yield of potential stem cells in cord blood

Human Dental Pulp Stem Cells: recent findings and current research

Abstract Prevalence of neurodegenerative diseases, most of which are life threatening and incurable, is an increasing clinical problem. To date, studies have demonstrated a superior proliferation rate of dental pulp stem cells (DPSCs) compared to other mesenchymal stem cells in vitro. DPSCs has recently been recognized as a novel treatment strategy for neurodegenerative disease, due to their advanced potential for neurogenic differentiation. The oral cavity has been described as a promising source of dental pulp stem cells. DPSCs are widely used in regenerative dentistry holding alternative capacity for osteogenic differentiation and therefore new promises for tissue and whole tooth regeneration. Dental stem cell banking offers a plentiful source of stem cells representing great potential for cell reprogramming and thus cell therapy. Recently, the association of pulp stem cells with three – dimensional scaffold templates allows for building up naturally derived implants. This review introduces to unique properties of DPSCs and biological factors influencing mineralization, proliferation and differentiation of pulp stem cells. Latest research studies are compared in terms of effectiveness and limitations of techniques for the isolation of pulp stem cells, including the enzymatic digestion and the explant culture methods. Moreover, a short overview of most recent findings and clinical application of DPSCs is proffered including progress of current research and limitations still to be addressed in the nearest future. Finally, the article presents new advances in the area of regenerative dentistry and regenerative medicine, including three dimensional printing and three dimensional analysis, emerged to deepen studies under procedures to replace the non patient specific artificial implants. Running title: DPSCs - review