Source Of Adult Stem Cells Research Articles

A Simple and Effective Mechanical Method for Adipose-Derived Stromal Vascular Fraction Isolation.

Over the last decades, there has been ongoing and evolving research concerning regenerative medicine, specifically, stem cells. The most common source of adult mesenchymal stem cells (MSCs) remains the adipose tissue and the easiest way to obtain such tissue is lipoaspirate. The fatty tissue obtained can be processed either in an enzymatic way, which is time-consuming and expensiveand carries several dangers for the viability of the stem cells included, or with mechanical means which are fast, inexpensive, yield enough viable cells, and can be readily used for autologous transplantation in one-stage procedures. Herein, we demonstrate our non-enzymatic method for obtaining adipose-derived stromal vascular fraction comprising MSCs. The stromal vascular fraction was isolated via centrifugation, and the characteristics and numbers of the cells isolated have been tested with flow cytometry assay, cell culture, and differentiation. Over 91% of viable MSCs were isolated using the mechanical method. The cells retained the ability to differentiate into osteocytes, adipocytes, and chondrocytes. The method presented is simple, requiring no special equipment, and yields a viable population of stem cells in large numbers. These cells can be readily used in several operations (orthopedic, dentistry, fistulas, etc.) making feasible "one-stage" procedures, thus proving their benefits for the patient and the health care system.

Therapeutic Potential in Wound Healing of Allogeneic Use of Equine Umbilical Cord Mesenchymal Stem Cells.

Wound healing after skin injury is a complex process, particularly in equines where leg wounds are prevalent and their repair is complicated due to the anatomical characteristics. Conventional treatments are not effective enough. The umbilical cord offers an unlimited source of adult mesenchymal stem cells (ucMSCs) from Wharton's jelly tissue. The present study aims to demonstrate the safety and therapeutic potential of the allogeneic use of equine ucMSCs (e-ucMSCs) in the healing of severe equine leg wounds. The methods employed were the isolation, culture and expansion of e-ucMSCs. Flow cytometry and a PCR assay were used for cell characterization. This study included an immunomodulation assay, a murine pre-clinical trial and the first phase of an equine clinical trial. Our results showed that e-ucMSCs express a functional HLA-G homolog, EQMHCB2. In the immunomodulation assay, the e-ucMSCs inhibited the proliferation of activated equine peripheral blood mononuclear cells (e-PBMCs). In the murine pre-clinical trial, e-ucMSCs reduced healing time by 50%. In the equine clinical trial, the injection of e-ucMSCs into severe leg lesions improved the closure time and quality of the tissues involved, regenerating them without fibrous tissue scar formation. In conclusion, the results of this study suggest that e-ucMSCs can be used allogeneically for wound healing by creating a tolerogenic environment.

Isolation of Human Adipose-Derived Stem Cells.

Human adipose-derived stromal/stem cells (hASCs) are a promising source of adult stem cells used in numerous applications in regenerative medicine. We present the protocols from our laboratory for isolating and expanding hASCs. The isolation of hASCs involves the enzymatic digestion of adipose tissue and subsequent culturing of the isolated cells.

Dental Pulp-Derived Mesenchymal Stem Cells Increase Axon Numbers in Mental Nerve Repair.

The mental nerve, the extended part of the inferior alveolar nerve, is often injured during dentoalveolar, orthognathic, or tumor surgery. Numerous therapeutic interventions, including surgery and pharmacotherapy, have been used to enhance the recovery of nerve injuries. Dental pulp stem cells (DPSCs) represent an easily accessible source of adult stem cells that can be isolated from the pulp of extracted teeth. This study evaluated the effect of DPSCs on the regeneration of the mental nerve injury model of rabbits. In this presented study, DPSCs were cultured and cell characterizations were performed by using flow cytometry and immunostainings. Bilateral mental nerve injury models of rabbits were created. In the control group (n = 10), saline was applied, and in the study group (n = 10), 2 × 106 DPSCs were applied to the repaired nerve areas. After 3 weeks, animals were killed and histological examination was obtained by using Masson's trichrome staining. An unpaired Student's t test was used when comparing the groups. Differences were considered to be statistically significant at P values of less than 0.05. The DPSCs demonstrated a homogeneous population of mesenchymal stromal cells which expressed cluster of differentiation CD44, CD73, CD90, and CD105 and lack of CD34, CD45, and HLA-DR. Our finding clearly demonstrated that a lower number of cross-sectioned axons were founded in the control group (60.18 ± 2.52) compared to the study group (72.96 ± 2.43) (p = 0.00). DPSCs promote mental nerve axonal regeneration. These results suggest that DPSCs provide an important accessible source of adult stem cells for mental nerve regeneration.

A Novel Benchtop Device for Efficient and Simple Purification of Cytokines, Growth Factors and Stem Cells from Adipose Tissue.

Lipoaspirates represent a source of adult stem cells, cytokines, and growth factors of adipocyte origin with immunomodulation and regenerative medicine potential. However, rapid and simple protocols for their purification using self-contained devices that can be deployed at the points of care are lacking. Here, we characterize and benchmark a straightforward mechanical dissociation procedure to collect mesenchymal stem cells (MSCs) and soluble fractions from lipoaspirates. IStemRewind, a benchtop self-contained cell purification device, allowed a one-procedure purification of cells and soluble material from lipoaspirates with minimal manipulation. The recovered cellular fraction contained CD73+, CD90+, CD105+, CD10+ and CD13+ MSCs. These markers were comparably expressed on MSCs isolated using IstemRewind or classic enzymatic dissociation procedures, apart from CD73+ MSCs, which were even more abundant in IStemRewind isolates. IstemRewind-purified MSCs retained viability and differentiation into adipocytes and osteocytes, even after a freezing-thawing cycle. Levels of IL4, IL10, bFGF and VEGF were higher compared to the pro-inflammatory cytokines TNFα, IL1β and IL6 in the IStemRewind-isolated liquid fraction. In sum, IStemRewind can be useful for straightforward, rapid, and efficient isolation of MSCs and immunomodulatory soluble factors from lipoaspirates, opening the possibility to directly isolate and employ them at the point-of-care.

149 - Outcomes of Cord Blood Vs Adult Stem Cell Sources in Patients with Mutated TP53 Myeloid Malignancies

149 - Outcomes of Cord Blood Vs Adult Stem Cell Sources in Patients with Mutated TP53 Myeloid Malignancies

Effects of green light-emitting diode irradiation on neural differentiation of human umbilical cord matrix-derived mesenchymal cells; Involvement of MAPK pathway

Alteration of the proliferation rate and differentiation of mesenchymal stem cells (MSCs) into various lineages, including neural cells, by light emitting diodes (LEDs) irradiation has received considerable attention in recent years. Human umbilical cord matrix-derived mesenchymal cells (hUCMs) are an accessible source of adult stem cells with appropriate characteristics that make them ideal tools for stem cell researches, cell therapy procedures and regenerative medicine. The aim of the present study was to investigate the effects of green LED irradiation, retinoic acid (RA) and their combination on the differentiation of hUCMs into neural lineage as well as the mechanisms involved. Exposure of hUCMs to green LED (530 nm, 1.59 J/cm2) with or without retinoic acid (RA) treatment, significantly increased the expression of specific genes including nestin, β-tubulin III, MAP2 and Olig2. In addition, immunohistochemical analysis confirmed expression of specific neural-related proteins including MAP2, GFAP and Olig2 in irradiated cells. ROS generation significantly increased following green light irradiation which in turn has activated the MAPK signaling pathway, resulting in the differentiation of hUCMs into neurons and glial cells, confirmed by western blot analysis of MAPK-related pathway. Taken together, our results suggest that the green LED irradiation, alone and in combination with RA, via ERK 1/2, JNK and p38 phosphorylation improves differentiation of hUCMs into neural lineage. Other mechanisms and inducers to enhance differentiation phenomena in vitro and in vivo should be investigated to determine the most appropriate strategy for therapeutic purposes.

Berberine Encapsulated in Exosomes Derived from Platelet-Rich Plasma Promotes Chondrogenic Differentiation of the Bone Marrow Mesenchymal Stem Cells via the Wnt/β-Catenin Pathway.

Cartilage regenerative medicine, wherein the stem cells from adults exert a crucial role, has high potential in the treatment of defective articular cartilage. Recently, Bone marrow mesenchymal stem cells (BMSCs) are being increasingly recognized as an alternative source of adult stem cells, which are capable of differentiating into several cell types (e.g., adipocytes, chondrocytes, and osteoblasts). However, their proliferative properties and tendency to dedifferentiate restrict their use in clinical settings. Recently, a possible bioactive material PRP-exos (exosomes derived from platelet-rich plasma), has emerged, which can effectively facilitate the differentiation and proliferation of cells. Recent studies have reported that berberine (Ber), known to have anti-inflammatory properties, plays a role in osteogenesis. Since biological molecules are used in combinations, we attempted to assess the effect of Exos-Ber (PRP-exos in combination with Ber) on the chondrogenic differentiation of BMSCs in vitro. In this study, Exos-Ber was observed to promote the proliferation of BMSCs and cause their chondrogenic differentiation in vitro. Additionally, Exos-Ber could promote the migration of BMSCs and increase the protein expression of the chondrogenic genes (Collagen II, SOX9, Aggrecan). After treatment with Exos-Ber, significant induction of β-catenin expression was observed, which could be repressed successfully by adding β-catenin inhibitor XAV-939. Interestingly, the repression of the Wnt/β-catenin axis also resulted in reduced gene expression levels of Collagen II, SOX9, and Aggrecan. These observations indicated that Exos-Ber facilitated the differentiation of chondrogenic BMSCs by modulating the Wnt/β-catenin axis, which offers innovative insights into the reconstruction of cartilage.

Knowledge, Awareness and Perception of Dental Stem Cell and Their Applications in Regenerative Medicine Among Professional Groups

Introduction:Dental stem cell research has become an important field in regenerative medicine. Nevertheless, the depth of knowledge and awareness of dental stem cells remain vague among the professional groups in Malaysia. The objective of this study is to assess the level of knowledge and awareness of professional groups in Malaysia on dental stem cells and their applications in regenerative medicine.Methods:A descriptive cross-sectional survey was implemented using online and hard-copy questionnaires that was distributed among medical doctors, scientists, and dentists to assess the level of knowledge and awareness (n=157). The questionnaire consists of 3 sections consisting of demographics, knowledge assessment and perception among the professional groups. The data were analysed using SPSS analysis.Results:Results showed that professional groups in Malaysia were knowledgeable and aware of dental stem cells and their application. Majority of the respondents were dentists (41.91%), followed by medical doctors (35.29%) and scientists (22.79%). 50.74% of respondents were unsure about the similar characteristics between dental stem cells (DSCs) and bone marrow mesenchymal stem cells (BMMSCs). Most respondents were aware of the derivation of DSCs from pulp and periodontal ligament (PDL) tissues (77.21%) and that DSCs are considered an adult stem cell source (51.11%). Most respondents were familiar with the characteristics of DSCs. They were aware that DSCs have the unique ability to self-renew and differentiate into trilineage differentiation (69.12%). They agreed that DSCs are an excellent cell resource for therapeutic approaches to neural repair and regeneration (63.24%). 63.24% of the respondents demonstrated a positive outcome for dental stem cell research. 59.56% of them would be willing to donate their extracted teeth for research. About 56.62% of respondents would conserve their teeth in stem cell banks and 52.94% are interested in investing in the stem cell industry.Conclusion:A high level of perception and awareness about dental stem cells and its application was noted among professional groups in Malaysia. They are supportive and willing to invest for dental stem cell research.

Stem Cells Era in Maxillofacial Surgery

Stem cells provide a new era for oral and maxillofacial surgery and reconstruction. Stem cells are colonlogic cells that have the ability to renew themselves and form various predecessors. They are found in common tissues used for wound healing and recovery after injuries. Large numbers of these cells are known to be instantly accessible; no immunological denial or rejection; do not have host disease against graft; Tumorigenesis do not occur; but there are some small ethical problems; The predictable differentiation potential for tissue separation and integration is not surprising. There are two basic types of stem cells, depending on their origin: • Embryonic Stem Cells (ESCs): ESCs are initiated by a fetus that has been fertilized in an in vitro fertilization clinic (100,200 organized cell blastocysts). They are called a pluripotent cell population and are characterized by the ability to maintain the karyotype. ESCs can be divided into the three embryonic cotyledons (ectoderm, endoderm, and mesoderm); and in a culture; they form in colonies. These important highlights make ESCs affordable for future use in regenerative medicine and pharmaceuticals. Still; ethical discussions about the use of human embryos cannot be decided entirely; because in addition to scientific and practical questions, it is also dealt with through religious laws and philosophical reflections on the nature of human life. • Adult stem cells: Adult stem cells are found in fetal and postnatal tissues; They are more specialized (multipotent) cells that participate in the repair and restoration of tissues, as well as in the maturation and aging processes. There are numerous sources of adult stem cells such as bone marrow (BMMSC); Adipose tissue (ATMSCs); Liver; Umbilical cord; and muscles. The intraoral sources are the dental pulp; The periodontal ligaments can also be a source of adult stem cells. Fat and bone marrow stem cells are successful and are produced in the context of restorative work and aesthetics. However, ATMSCs have been less easily obtained without harmful effects on patients. Adipose tissue contains a population of mesenchymal stem cells that can be isolated and differentiated into various cell lines, including osteocytes; Fat cells; and myocytes depending on the culture conditions. These cells are called adipose derived stem cells (ASC). The question arises here is: what do stem cells do? And the answer is so simple when you know that 10 million cells in your body die every minute every day and your own stem cells replace them so you can continue living. Both undifferentiated ATMSC and BMMSC have osteogenic; chondrogenic and lipogenic differentiation potential. Both produce markers that help repair the deformity and have the same morphology; spindle shaped cells. Both appeared to work positively when used in reconstructive treatments. There are some contrasts between them; but the main imperative is; the ease of collection and the lower morbidity on the part of the donor. These two variables were extremely critical and favor the choice between fat and bone marrow for repair. ATMSCs and BMMSCs may be suitable replacements for the traditional intrusive method in the future. A better understanding of stem cell types, different platforms, and the influence of developmental components on the tissue design of different scaffolds, and how growth factors affect the tissue engineering pathway, opens opportunities to monitor recovery of bones and soft defects in the future, thus providing an alternative and innovative treatment for patients with soft / hard tissue defects.

Chemically Defined Conditions Mediate an Efficient Induction of Dental Pulp Pluripotent-Like Stem Cells into Hepatocyte-Like Cells

Liver diseases are major causes of morbidity and mortality. Dental pulp pluripotent-like stem cells (DPPSCs) are of a considerable promise in tissue engineering and regenerative medicine as a new source of tissue-specific cells; therefore, this study is aimed at demonstrating their ability to generate functional hepatocyte-like cells in vitro. Cells were differentiated on a collagen scaffold in serum-free media supplemented with growth factors and cytokines to recapitulate liver development. At day 5, the differentiated DPPSC cells expressed the endodermal markers FOXA1 and FOXA2. Then, the cells were derived into the hepatic lineage generating hepatocyte-like cells. In addition to the associated morphological changes, the cells expressed the hepatic genes HNF6 and AFP. The terminally differentiated hepatocyte-like cells expressed the liver functional proteins albumin and CYP3A4. In this study, we report an efficient serum-free protocol to differentiate DPPSCs into functional hepatocyte-like cells. Our approach promotes the use of DPPSCs as a new source of adult stem cells for prospective use in liver regenerative medicine.

Neonatal Porcine Germ Cells Dedifferentiate and Display Osteogenic and Pluripotency Properties.

Gonocytes are progenitors of spermatogonial stem cells in the neonatal testis. We have previously shown that upon culturing, neonatal porcine gonocytes and their colonies express germ cell and pluripotency markers. The objectives of present study were to investigate in vitro trans-differentiation potential of porcine gonocytes and their colonies into cells from three germinal layers, and to assess pluripotency of cultured gonocytes/colonies in vivo. For osteogenic and tri-lineage differentiation, cells were incubated in regular culture media for 14 and 28 days, respectively. Cells were cultured for an additional 14 days for osteogenic differentiation or 7 days for differentiation into derivates of the three germinal layers. Osteogenic differentiation of cells and colonies was verified by Alizarin Red S staining and tri-lineage differentiation was confirmed using immunofluorescence and gene expression analyses. Furthermore, upon implantation into recipient mice, the cultured cells/colonies developed teratomas expressing markers of all three germinal layers. Successful osteogenic differentiation from porcine germ cells has important implications for bone regeneration and matrix formation studies. Hence, gonocytes emerge as a promising source of adult pluripotent stem cells due to the ability to differentiate into all germinal layers without typical biosafety risks associated with viral vectors or ethical implications.

Endometrial regenerative cells with galectin-9 high-expression attenuate experimental autoimmune hepatitis

BackgroundAutoimmune hepatitis (AIH) is a T cell-mediated immune disease that activates abnormally against hepatic antigens. We have previously reported that endometrial regenerative cells (ERCs) were a novel source of adult stem cells, which exhibiting with powerful immunomodulatory effects. Galectin-9 (Gal-9) is expressed in ERCs and plays an important role in regulating T cell response. This study aims to explore the role of ERCs in attenuation of AIH and to determine the potential mechanism of Gal-9 in ERC-mediated immune regulation.MethodsERCs were obtained from menstrual blood of healthy female volunteers. In vitro, ERCs were transfected with lentivirus vectors carrying LGALS9 gene and encoding green fluoresce protein (GFP-Gal-9-LVs) at a MOI 50, Gal-9 expression in ERCs was detected by ELISA and Q-PCR. CD4+ T cells isolated from C57BL/6 mouse spleen were co-cultured with ERCs. The proliferation of CD4+ T cells was detected by CCK-8 kit and the level of Lck/zap-70/LAT protein was measured by western blot. Furthermore, AIH was induced by ConA in C57BL/6 mice which were randomly assigned to untreated, unmodified ERC-treated and Gal-9 high-expressing ERC-treated groups. Histopathological score, liver function, CD4+/CD8+ cell infiltration in liver tissues, the proportion of immune cells in the spleen and liver, and ERC tracking were performed accordingly to assess the progression degree of AIH.ResultsAfter transfecting with GFP-Gal-9-LVs, Gal-9 expression in ERCs was significantly increased. Additionally, Gal-9 high-expressing ERCs effectively inhibited CD4+ T cell proliferation and downregulated CD4+ T cell active related proteins p-Lck/p-ZAP70/p-LAT in vitro. Furthermore, treatment with Gal-9 high-expressing ERCs restored liver function, ameliorated liver pathological damage, inhibit CD4+ and CD8+ T cell proliferation and suppress Th1 and Th17 cell response in the hepatitis mice. In addition, Gal-9 high-expressing ERCs further markedly enhanced the level of IL-10 but reduced the levels of IFN-γ, TNF-α, and IL-4 in mouse sera and liver. Cell tracking also showed that ERCs could migrate to the damaged liver organs.ConclusionsThe results suggested that Gal-9 was an essential modulator, which was required by ERCs in regulating T cell response and attenuating ConA-induced experimental hepatitis. And also, it provides a novel idea for the clinical treatment of AIH.

Therapeutic Potential of Human Nasal Inferior Turbinate-Derived Stem Cells: Microarray Analysis of Multilineage Differentiation

Introduction: Human nasal inferior turbinate-derived stem cells (hNTSCs) are attractive sources of adult stem cells for medical application because they can be easily obtained and cultivated with a highly proliferative capacity. The ability of hNTSCs to differentiate into chondrocytes, osteocytes, and neural cells makes them potential replacement therapeutic candidates in intractable disease. Nevertheless, detailed expression pattern of genes associated with trilineage differentiation (osteogenesis, chondrogenesis, and neurogenesis) in hNTSCs has not been revealed yet. Methods: In this study, we aimed to evaluate gene expression patterns of various transcription factors and marker genes associated with a particular lineage (osteogenesis, chondrogenesis, and neurogenesis) of differentiation of hNTSCs by DNA microarrays. Results: In microarrays, 36 transcripts such as E2F transcription factor 1, activating transcription factor 5, and AKR1B10 were upregulated and 36 transcripts such as CA9, PPFIA4, HAS2, and COL4A4 were downregulated in osteogenically differentiated hNTSCs. In chondrogenically differentiated hNTSCs, 3 transcripts (NUDT14, CPA4, and heparin-binding epidermal growth factor-like growth factor) were upregulated and 82 transcripts such as PTGS1, CLEC2D, and TET1 were downregulated. In neurally differentiated hNTSCs, 61 transcripts such as insulin-like growth factor-binding protein-1, nerve growth factor receptor, FGF1, OLFML1, and EPGN were upregulated and 98 transcripts such as ACAN, RUNX2, and C21orf96 were downregulated. In gene ontology (GO) analysis, cell signal-related GO terms were highly expressed. By contrast, catalysis GO terms and GO terms related to oxidoreductase were overrepresented in chondrogenically differentiated hNTSCs and osteogenically differentiated hNTSCs, respectively. Conclusion: Considering overall results, hNTSCs-specific genetic information may promote further studies on intracellular mechanisms defining key features of these cells.

Construction of a Blood Vessel Wall with Adipose-Derived Stem Cell Sheets In Vitro

Adipose-derived stem cells (ASCs) are a reliable and easily accessible source of adult stem cells. Thus, ASCs are increasingly used as seed cells in tissue engineering. This report investigated the possibility of engineering a large vessel using unwoven polyglycolic acid (PGA) fibers covered with ASC sheets treated with transforming growth factor-β1 (TGF-β1) and platelet-derived growth factor-BB (PDGF-BB). After coiling around a silicone tube, the complex was secured by surgical sutures and cultured In Vitro for six weeks. Gross views showed that glossy and smooth tubular tissue was engineered. Histological examination revealed regularly distributed cells and a few collagenous and elastic fibers in the differentiated group. Cells were randomly distributed, and disorganized collagenous and small elastic fibers were detected in the control group. Our results indicate the promising potential of ASC sheets and PGA fibers for the construction of large vessel walls.

Potential application of human neural crest-derived nasal turbinate stem cells for the treatment of neuropathology and impaired cognition in models of Alzheimer\u2019s disease

BackgroundStem cell transplantation is a fascinating therapeutic approach for the treatment of many neurodegenerative disorders; however, clinical trials using stem cells have not been as effective as expected based on preclinical studies. The aim of this study is to validate the hypothesis that human neural crest-derived nasal turbinate stem cells (hNTSCs) are a clinically promising therapeutic source of adult stem cells for the treatment of Alzheimer’s disease (AD).MethodshNTSCs were evaluated in comparison with human bone marrow-derived mesenchymal stem cells (hBM-MSCs) according to the effect of transplantation on AD pathology, including PET/CT neuroimaging, immune status indicated by microglial numbers and autophagic capacity, neuronal survival, and cognition, in a 5 × FAD transgenic mouse model of AD.ResultsWe demonstrated that hNTSCs showed a high proliferative capacity and great neurogenic properties in vitro. Compared with hBM-MSC transplantation, hNTSC transplantation markedly reduced Aβ42 levels and plaque formation in the brains of the 5 × FAD transgenic AD mice on neuroimaging, concomitant with increased survival of hippocampal and cortex neurons. Moreover, hNTSCs strongly modulated immune status by reducing the number of microglia and the expression of the inflammatory cytokine IL-6 and upregulating autophagic capacity at 7 weeks after transplantation in AD models. Notably, compared with transplantation of hBM-MSCs, transplantation of hNTSCs significantly enhanced performance on the Morris water maze, with an increased level of TIMP2, which is necessary for spatial memory in young mice and neurons; this difference could be explained by the high engraftment of hNTSCs after transplantation.ConclusionThe reliable evidence provided by these findings reveals a promising therapeutic effect of hNTSCs and indicates a step forward the clinical application of hNTSCs in patients with AD.

Assessment of Pregnant Women knowledge and attitude towards Umbilical Cord Banking and Stem Cell.

Background: Nowadays, the collection of stem cells from the blood of the umbilical cord is one of the most important topics of health sciences and new technology. However, cord blood is now considered a rich source of adult stem cells that can be used to treat many conditions and diseases. It is still a controversial topic. The current study aims to assess pregnant women's knowledge and attitude toward umbilical cord banking and stem cell. Research design: Descriptive exploratory research design was adopted to achieve the aim of the study. Setting: The study was conducted at the obstetrics &gynecology outpatient clinic at Beni-Suef University hospital in Egypt. Subjects: A convenient sample of 200 pregnant women was recruited. Tools of data collection: three tools were used for this study (1) Maternal structured interviewing questionnaire and (2) knowledge assessment questionnaire and (3) attitude assessment questionnaire. Results: It was found that 42.5% had an average level of knowledge regarding cord blood banking and stem cell with the negative attitude (67%) with no statistically significant difference (p >0.05). Besides, the sources of information were social media followed by nurses/ midwives. Conclusion: The present study concluded that despite nearly half of the pregnant women had average level knowledge but the majority of them had a negative attitude toward umbilical cord blood banking and stem cell. Recommendations: adequately planned in-service training programs related to cord blood collection and stem cells established to distribute among the pregnant women who attending the antenatal clinics.

Dental stem cell banking: Techniques and protocols.

Dental tissue-derived stem cells (DSCs) provide an easy, accessible, relatively noninvasive promising source of adult stem cells (ASCs), which brought encouraging prospective for their clinical applications. DSCs provide a perfect opportunity to apply for a patient's own ASC, which poses a low risk of immune rejection. However, problems associated with the long-term culture of stem cells, including loss of proliferation and differentiation capacities, senescence, genetic instability, and the possibility of microbial contamination, make cell banking necessary. With the rapid development of advanced cryopreservation technology, various international DSC banks have been established for both research and clinical applications around the world. However, few studies have been published that provide step-by-step guidance on DSCs isolation and banking methods. The purpose of this review is to present protocols and technical details for all steps of cryopreserved DSCs, from donor selection, isolation, cryopreservation, to characterization and quality control. Here, the emphasis is on presenting practical principles in accordance with the available valid guidelines.

Evaluation of Culture Morphology of Neuronally Transdifferentiated Wharton’s Jelly Derived Mesenchymal Stem Cells

Background: The prospect of mesenchymal stem cells (MSCs) as an adult stem cell source for neuronal tissue regeneration via their ability to differentiate into neurons has generated considerable excitement in regenerative cell therapy.Methods: In this study, we isolated ovine Wharton’s jelly derived MSCs and expanded in vitro in adherent culture. After the characterisation of MSCs using specific markers, we analysed the culture morphology of MSCs differentiated into neurons by a two-step chemical-based induction protocols involving a pre-induction step and a direct one step chemical-based induction protocol. Morphological changes after induction were evaluated.Result: In both the methods, after neuronal induction, the cells displayed phenotypic characteristic of neurons and comparatively less cytotoxicity was observed in the direct induction method. This study confirmed the possibility of generating neuron like cells from ovine WJ-MSCs and thereby exploring the potential of MSCs as therapeutic tool for treating neurological disorders in Veterinary Medicine.

Decreased Inhibition of Proliferation and Induction of Apoptosis in Breast Cancer Cell Lines (T47D and MCF7) from Treatment with Conditioned Medium Derived from Hypoxia-Treated Wharton's Jelly MSCs Compared with Normoxia-Treated MSCs.

Background: Mesenchymal stem cells (MSCs) are an appealing source of adult stem cells for cell therapy due to the high rate of proliferation, self-renewal capability, and applicable therapy. Wharton’s jelly (WJ), the main component of the umbilical cord extracellular matrix, comprises multipotent stem cells with a high proliferation rate and self-renewal capability and has anti-cancer properties. MSCs have been reported to secrete a variety of cytokines that have a cytotoxic effect in various cancers. Oxygen tension affects MSCs proliferation, cytokines level but no in surface markers expression, MSCs’ differentiation. We explored the cytotoxic effect and inducing apoptosis of Wharton’s jelly derived mesenchymal stem cells (WJMSCs) secretions from normoxic WJMSCs (WJMSCs-norCM) (CM: conditioned medium) and hypoxic WJMSCs (WJMSCs-hypoCM) in breast cancer cell lines (T47D and MCF7). Materials and Methods: Cytotoxic activity was determined using the MTS assay. RT-PCR was performed to measure the expression of apoptosis-inducing genes, specifically P53, BAX, and CASP9, and the antiapoptotic gene BCL-2. Results: WJMSCs-norCM and WJMSCs-hypoCM were potent inhibitors of the proliferation in both cell lines. WJMSCs-norCM had more anticancer activity in T47D and MCF7. The IC50 value of WJMSCs-norCM on MCF7 was 42.34%, and on T47D was 42.36%. WJMSCs-norCM significantly induced the gene expression of apoptotic P53, BAX, and CASP9 and insignificantly decreased the antiapoptotic gene BCL-2 in both MCF7 and T47D cells. WJMSCs-CM has anticancer activity by inducing P53, BAX, and CASP9 apoptotic genes. Conclusion: WJMSCs-norCM has more anticancer activity than WJMSCs-hypoCM.