Umbilical Cord Wharton's Jelly Research Articles

Development of a simple selection protocol for optimizing the harvest of mesenchymal stem cells from explanted human umbilical cord Wharton’s jelly

Background: Human umbilical cord Wharton's jelly (hUCWJ) derived Mesenchymal stem cells (MSCs) is a non-controversial, easily available source of human tissue which has a close association with embryonic tissue. Explant culture technique of isolating MSCs do not require any proteolytic enzymes giving an edge when compared to enzymatic methods.
 Aims and Objective: This study was designed to develop a suitable protocol to optimize the number of primary cells by transferring the explants after a successful migration and before trypsinizing of MSCs from primary culture plate to new culture plates.
 Materials and Methods: For defining the isolated cells as MSCs, the cells were studied by colony forming assay, semiquantitative two steps reverse transcriptase polymerase chain reaction (RT-PCR) and International Society for Cellular Therapy (ISCT) criteria to determine the purity of the lineage.
 Results: The explant cultured cells fulfil the defining criteria by ISCT. Further, they form colony and RT-PCR confirmed the presence of thy-1, endoglin and β-actin. The incubated explant shows successful cells migration after transferring from one culture plate to another for five consecutive times. Further, it was seen that the migrated cells can be cultured for more than 18 passages. However, a reduction was seen in the migration of the cells after the 3rd transfer of the explants.
 Conclusion: This study found that the transferred explant has the potential to yield cells for 5 consecutive explant transfers. Further, this technique optimized the yield of primary cells and thereby reducing the quantity and frequency of sample collection, processing time and cost.

The healing effect of Wharton's jelly stem cells seeded on biological scaffold in chronic skin ulcers: A randomized clinical trial.

Chronic wound or nonhealing ulcer is essentially a wound that does not progress normally through the wound healing process. This study assessed the healing effect of umbilical cord Wharton's jelly stem cells seeded on biological scaffold in chronic skin ulcers. In a randomized clinical trial, five patients between 30 and 60years with chronic diabetic wounds were enrolled. To cover the wounds, acellular amniotic membrane seeded with Wharton's jelly mesenchymal stem cells (WJSCs) was used for 9days, every 3days with a follow-up of 1month. The percentage and time of wound healing and the size of wound were recorded for each patient. In treated patients, the wound healing time and wound size significantly decreased, and after 6 and 9days, the wound size significantly declined (P<0.002). As WJSCs seeded on amniotic membrane could significantly accelerate the healing effect in chronic diabetic wounds, they can be an alternative source in tissue engineering and repair of chronic ulcers.

Stability characteristic of cryopreserved human umbilical cord wharton's jelly–derived mesenchymal stromal cells

Background & Aim Mesenchymal stromal cells (MSCs) are multipotent mesodermal linage derived stem cells that can self–replicate and differentiate into all mesodermal, some of neuroectodermal and hepatic endodermal progenies. Umbilical cord derived Wharton's jelly MSCs (WJ–MSCs) are an attractive source for cell–based therapy regenerative medicine due to the stability in large scale expansion, non–tumor formation and low immunological rejection. Therefore, WJ–MSCs are a superior banking MSCs source for further clinical application. Methods, Results & Conclusion WJ–MSCs were isolated at a 100% successful rate and characterized by flow cytometry for CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1. Growth kinetics characteristics were investigated by population doublings (PD), clinical large–scale expansion and high viability maintenance. MSCs were assessed for differentiation potential to cartilage, adipose and bone. Flow cytometry analysis showed no differences in expression of CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1 among all cryopreserved WJ–MSCs for six years (2013–2018: n=30). All years of cryopreserved WJ–MSCs maintained high proliferative potential with no difference in all passages. Moreover, they were capable to differentiate into cartilage, adipose and bone structures as determined with histochemistry. WJ–MSCs have a high potential for stem cell banking from umbilical cord due to its noninvasive techniques of procurement, high successful isolation rate and low risk microorganism contamination. Furthermore, WJ–MSCs are suitable sources of stem cells for potential use in regenerative medicine in further clinical applications. Mesenchymal stromal cells (MSCs) are multipotent mesodermal linage derived stem cells that can self–replicate and differentiate into all mesodermal, some of neuroectodermal and hepatic endodermal progenies. Umbilical cord derived Wharton's jelly MSCs (WJ–MSCs) are an attractive source for cell–based therapy regenerative medicine due to the stability in large scale expansion, non–tumor formation and low immunological rejection. Therefore, WJ–MSCs are a superior banking MSCs source for further clinical application. WJ–MSCs were isolated at a 100% successful rate and characterized by flow cytometry for CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1. Growth kinetics characteristics were investigated by population doublings (PD), clinical large–scale expansion and high viability maintenance. MSCs were assessed for differentiation potential to cartilage, adipose and bone. Flow cytometry analysis showed no differences in expression of CD34, CD45, CD73, CD90, CD105, CD106 and STRO–1 among all cryopreserved WJ–MSCs for six years (2013–2018: n=30). All years of cryopreserved WJ–MSCs maintained high proliferative potential with no difference in all passages. Moreover, they were capable to differentiate into cartilage, adipose and bone structures as determined with histochemistry. WJ–MSCs have a high potential for stem cell banking from umbilical cord due to its noninvasive techniques of procurement, high successful isolation rate and low risk microorganism contamination. Furthermore, WJ–MSCs are suitable sources of stem cells for potential use in regenerative medicine in further clinical applications.

Histological, immunohistochemical, and genomic evaluation of excisional and diabetic wounds treated with human Wharton's jelly stem cells with and without a nanocarrier.

We showed in previous studies that human umbilical cord Wharton's jelly stem cells (hWJSCs) improved the healing rates of excisional and diabetic wounds in the mouse model. As an extension of those studies, we report here the more detailed quantitative histological, immunohistochemical, and genomic evaluation of biopsies from those excisional and diabetic wounds in an attempt to understand the mechanisms of the enhanced wound healing aided by hWJSCs. Bright-field microscopic observations and ImageJ software analysis on histological sections of the excisional and diabetic wound biopsies collected at different time points showed that the thickness of the epidermis and dermis, and positive picrosirius-red stained areas for collagen, were significantly greater in the presence of hWJSCs compared with controls (P < 0.05). Immunohistochemistry of the diabetic wound biopsies showed increased positive staining for the vascular endothelial marker CD31 and cell proliferation marker Ki67 in the presence of hWJSCs and its conditioned medium (hWJSC-CM). Quantitative real-time polymerase chain reaction showed upregulation of groups of genes involved in extracellular matrix regulation, collagen biosynthesis, angiogenesis, antifibrosis, granulation, and immunomodulation in the presence of hWJSCs. Taken together, the results demonstrated that hWJSCs and hWJSC-CM that contains the paracrine secretions of hWJSCs, enhance the healing of excisional and diabetic wounds via re-epithelialization, collagen deposition, angiogenesis, and immunomodulation. The inclusion of an Aloe vera-polycaprolactone (AV/PCL) nanocarrier did not significantly change the effect of the hWJSCs. However, the topical application of an AV/PCL nanocarrier impregnated with hWJSCs is convenient and less invasive than the administration of hWJSC injections into wounds.

Differentiation of umbilical cord Wharton's jelly derived mesenchymal stem cells into cardiomyocytes using 5-azacytidine

Objectives The objective of this article was to differentiate mesenchymal stem cells (MSCs) into cardiomyocytes. Background Cardiomyopathies resulted in permanent loss of cardiomyocytes as it had no ability of regeneration. This made MSCs a promising tool for cellular therapy because of their ability of self-renewal and multipotency. MSCs were considered ideal for cellular cardiomyoplasty as they can undergo full cardiogenic differentiation. Now it was believed that isolated cells from umbilical cord and expanded in vitro were a potential source of MSCs. Patients and methods An experimental study included 10 pregnant females due for delivery between October 2015 and April 2017. Umbilical cord samples and cord blood were collected from cesarean section patients, MSCs were cultured from umbilical cord tissue (Wharton's jelly). MSCs were subcultured in differentiating media containing azacytidine. Cardiomyocytes differentiation was detected by morphology of cardiomyocytes and immunophenotyping. Results MSCs were successfully isolated from 10 umbilical cord samples. MSCs showed positive expression of CD44 for umbilical cord mesenchymal stem cells (UCMSCs) mean ± SD (79.72 ± 5.85). It showed negative expression of CD34 for UCMSCs mean ± SD (1.08 ± 0.43). A significant statistical difference was found (P = 3.28 × 10−11) between MSCs and cardiomyocytes with respect to expression of troponin. A significant statistical correlation was seen (P = 0.001) between MSCs that showed positive expression of CD44 and cardiomyocytes. Conclusion By using azacytidine MSCs isolated from umbilical cord Wharton's jelly (UCWJ) can be differentiated into cardiomyocytes.

Hyperglycemia differentially affects proliferation, apoptosis, and BNIP3 and p53 mRNA expression of human umbilical cord Wharton's jelly cells from non-diabetic and diabetic pregnancies

Diabetes in pregnancy constitutes an unfavorable environment for embryonic and fetal development, where the child has a higher risk of perinatal morbidity and mortality, with high incidence of congenital malformations and predisposition to long-term metabolic diseases that increase with a hypercaloric diet. To analyze whether hyperglycemia differentially affects proliferation, apoptosis, and mRNA expression in cells from children of normoglycemic pregnancies (NGPs) and diabetes mellitus pregnancies (DMPs), we used umbilical cord Wharton jelly cells as a research model. Proliferation assays were performed to analyze growth and determine the doubling time, and the rate of apoptosis was determined by flow cytometry-annexin-V assays. AMPK, BNIP3, HIF1α, and p53 mRNA gene expression was assessed by semi-quantitative RT-PCR. We found that hyperglycemia decreased proliferation in a statistically significant manner in NGP cells treated with 40 mM D-glucose and in DMP cells treated with 30 and 40 mM D-glucose. Apoptosis increased in hyperglycemic conditions in NGP and DMP cells. mRNA expression of BNIP3 and p53 was significantly increased in cells from DMPs but not in cells from NGPs. We found evidence that maternal irregular metabolic conditions, like diabetes with hyperglycemia in culture, affect biological properties of fetal cells. These observations could be a constituent of fetal programming.

Epithelial In vitro Differentiation of Mesenchymal Stem Cells.

Mesenchymal stem cells or mesenchymal stromal cells (MSCs) are non-hematopoietic stromal cells that reside in many human organs and have been isolated from a variety of adult or fetal tissues such as adipose tissue, bone marrow and umbilical cord Wharton's jelly, among others. Because they are a heterogeneous population, International Society for Cellular Therapy has established 3 minimum criteria to characterize MSCs in vitro: i) adherence to plastic, ii) differentiation potential (osteogenic, chondrogenic and adipogenic lineages) and iii) expression of specific surface antigens (CD73+, CD90+, CD105+, CD34-, CD45-, CD11b-, CD14-, CD19-, CD79a-, HLA-DR-). Because of these characteristics, MSCs are useful for different applications and studies, most of them related with regenerative biomedicine. Epithelial in vitro differentiation of MSCs, for clinical use, is one of the main objectives in this field, due to, on the one hand, the difficulties to establish epithelial cell cultures and, on the other hand, the immunomodulatory capacity of MSCs that could increase the success of transplantation. According to this and the information compiled from bibliography, production of epithelial cells differentiated in vitro from MSCs is a complex procedure and a lot of techniques and culture media are necessary to explore. The objective of this review is to show the different methods of epithelial in vitro differentiation and remark the need to further study for being capable of establishing specific cell lines of epithelial cells differentiated from autogenic or allogenic MSCs.

Proliferative, Differentiative, and Immunological Characteristics of Chondro-Differentiated Mesenchymal Stromal Cells Derived from Rabbit Umbilical Cord Wharton's Jelly

Proliferative, Differentiative, and Immunological Characteristics of Chondro-Differentiated Mesenchymal Stromal Cells Derived from Rabbit Umbilical Cord Wharton's Jelly

Wharton's jelly-derived mesenchymal cells as a new source for the generation of microtissues for tissue engineering applications.

Microtissues (MT) are currently considered as a promising alternative for the fabrication of natural, 3D biomimetic functional units for the construction of bio-artificial substitutes by tissue engineering (TE). The aim of this study was to evaluate the possibility of generating mesenchymal cell-based MT using human umbilical cord Wharton's jelly stromal cells (WJSC-MT). MT were generated using agarose microchips and evaluated ex vivo during 28days. Fibroblasts MT (FIB-MT) were used as control. Morphometry, cell viability and metabolism, MT-formation process and ECM synthesis were assessed by phase-contrast microscopy, functional biochemical assays, and histological analyses. Morphometry revealed a time-course compaction process in both MT, but WJSC-MT resulted to be larger than FIB-MT in all days analyzed. Cell viability and functionality evaluation demonstrated that both MT were composed by viable and metabolically active cells, especially the WJSC during 4-21days ex vivo. Histology showed that WJSC acquired a peripheral pattern and synthesized an extracellular matrix-rich core over the time, what differed from the homogeneous pattern observed in FIB-MT. This study demonstrates the possibility of using WJSC to create MT containing viable and functional cells and abundant extracellular matrix. We hypothesize that WJSC-MT could be a promising alternative in TE protocols. However, future cell differentiation and in vivo studies are still needed to demonstrate the potential usefulness of WJSC-MT in regenerative medicine.

Functional organic cation transporters mediate osteogenic response to metformin in human umbilical cord mesenchymal stromal cells

BackgroundCompelling evidence indicates that metformin, a low-cost and safe orally administered biguanide prescribed to millions of type 2 diabetics worldwide, induces the osteoblastic differentiation of mesenchymal stromal cells (MSCs) through the 5' adenosine monophosphate (AMP)-activated protein kinase (AMPK) pathway. As a highly hydrophilic cationic compound, metformin uptake is facilitated by cell membrane organic cation transporters (OCTs) of the solute carrier 22A gene family. We hypothesized that to effectively enhance osteogenic differentiation, and ultimately bone regeneration, metformin must gain access into functional OCT-expressing MSCs. MethodsData was obtained through immunoblotting, cellular uptake, mineralization and gene expression assays. ResultsWe demonstrate for the first time that functional OCTs are expressed in human-derived MSCs from umbilical cord Wharton's jelly, an inexhaustible source of nonembryonic MSCs with proven osteogenic potential. A clinically relevant concentration of metformin led to AMPK activation, enhanced mineralized nodule formation and increased expression of the osteogenic transcription factor Runt-related transcription factor 2 (RUNX2). Indeed, targeting OCT function through pharmacological and genetic approaches markedly blunted these responses. ConclusionsOur findings indicate that functional OCT expression in UC-MSCs is a biological prerequisite that facilitates the intracellular uptake of metformin to induce an osteogenic effect. Future pre-clinical studies are warranted to investigate whether the expression of functional OCTs may serve as a potential biomarker to predict osteogenic responses to metformin.

Chemically Defined Conditions Mediate an Efficient Induction of Mesodermal Lineage from Human Umbilical Cord- and Bone Marrow- Mesenchymal Stem Cells and Dental Pulp Pluripotent-Like Stem Cells.

The human umbilical cord Wharton's Jelly- and the bone marrow- mesenchymal stem cells (WJ-MSCs and BM-MSCs, respectively) and the newly identified dental pulp pluripotent-like stem cells (DPPSCs) are new sources for stem cells with prospective use in cell regeneration and therapy. These cells are self-renewable, can be differentiated into several lineages, and can potentiate the immune responses. We hypothesized that three-dimensional (3D) culture conditions and directed differentiation using specific signaling regulators will enhance an efficient generation of mesoderm (MD) lineage independent from the origin or source of the stem cells. For a period of 3-days, cell aggregates were generated in a serum-free media containing ascorbic acid, retinoic acid, and keratinocyte growth factor; sonic hedgehog and bone morphogenic protein-4 signaling were inhibited using small molecules. In all cell types used, the biochemical and molecular analysis revealed a time course-dependent induction of the mesodermal, but not endodermal or ectodermal makers. In this study, we utilized a novel and efficient serum-free protocol to differentiate WJ-MSCs, BM-MSCs, and DPPSCs into MD-cells. Successful development of an efficient differentiation protocol can further be utilized and expanded on to obtain MD- derivative cell lineages.

Human umbilical cord Wharton's jelly mesenchymal stem cells combined with an acellular cartilage extracellular matrix scaffold improve cartilage repair compared with microfracture in a caprine model

As a novel and promising seed cell, human umbilical cord Wharton's jelly mesenchymal stem cells (hWJMSCs) are widely applied in tissue engineering. However, whether hWJMSCs can better repair and regenerate the articular cartilage in big animals than microfracture (MF, a predominant clinical treatment strategy for damaged cartilage) is unclear. Evaluation of the validity, and safety of hWJMSCs in a caprine model with a full-thickness femoral condyle articular cartilage defect, compared with MF is required. After cultivation and identification, hWJMSCs were seeded in an acellular cartilage extracellular matrix (ACECM)-oriented scaffold to construct cell-scaffold complex. Six goats with full-thickness femoral condyle articular cartilage defects were randomized to MF (microfracture group, MFG) and cell-scaffold complexes (experimental group, EG). At 2 and 4 weeks, joint fluid was used to assess immuno-inflammatory responses. At 6 and 9 months, all goats were euthanized for assessment of morphology, and magnetic resonance imaging (MRI), histology staining, and evaluation of the elasticity modulus and glycosaminoglycan (GAG) contents of the repaired regions. There were no significant differences between the two groups in immuno-inflammatory parameters. MRI demonstrated higher-quality cartilage and complete subchondral bone at defect sites in the EG at 9 months. Histological staining showed that extracellular cartilage, cartilage lacuna and collagen type II levels were higher in the EG compared to the MFG, while the EG exhibited a higher elasticity modulus. The hWJMSCs-ACECM scaffold complex achieved better quality repair and regeneration of hyaline cartilage without cartilage-inducing factor, while retaining the structure and functional integrity of the subchondral bone, compared with MF.

Monitoring wound healing of burn in rat model using human Wharton's jelly mesenchymal stem cells containing cGFP integrated by lentiviral vectors.

Objective(s):Human Wharton’s Jelly mesenchymal stem cells (hWMSCs) are undifferentiated cells commonly used in regenerative medicine. The aim of this study was to develop a reliable tool for tracking hWMSCs when utilized as therapeutics in burnt disorders and also to optimize the cell-based treatment procedure.Materials and Methods:The hWMSCs were first isolated from fresh umbilical cord Wharton’s jelly and cultured. The 293LTV cell line was transfected by cGFP containing lentiviral vector and the helper plasmids for production of the viral particle. The viral particles were collected to transduce the hWMSCs. The transduced cells were finally selected based on resistance to puromycin. The burned rats (n=24) were treated with cGFP expressing hWMSCs using the cell spray method, with the cells being tracked 7, 14 and 21 days later. The rats were sacrificed 7, 14 and 21 days following treatment and paraffin embedded sections prepared from the burned area for downstream pathological analyses.Results:The lentiviral particles carrying the cGFP gene were generated and the hWMSCs were transduced. The cGFP-expressing hWMSCs were detected in the burned tissue and the burned injuries were improved dramatically as compared to control.Conclusion:Because of the establishment of stably transduced cGFP expressing cells and the ability to detect cGFP for a relatively long-time interval, the method was found to be quite efficient for the purpose of cell tracking. The combination of hWMSC-based cell therapy and sterile Gauze Vaseline (GV) as covering was proven much more efficient than the traditional methods based on GV alone.

Cartilage Repair in the Knee Using Umbilical Cord Wharton's Jelly–Derived Mesenchymal Stem Cells Embedded Onto Collagen Scaffolding and Implanted Under Dry Arthroscopy

Cell-based cartilage repair procedures are becoming more widely available and have shown promising potential to treat a wide range of cartilage lesion types and sizes, particularly in the knee joint. More recently, techniques have evolved from 2-step techniques that use autologous chondrocyte expansion to 1-step techniques that make use of mesenchymal stem cells (MSCs) embedded onto biocompatible scaffolding. Our 1-step technique has been further developed to provide cell-based cartilage repair using MSCs that have the potential to be used in an off-the-shelf manner, without the need for autologous tissue harvest. Precursor MSCs can be isolated in abundance from the Wharton's jelly of umbilical cord tissue. These cells have been shown to have the desired capacity for proliferation, differentiation, and release of trophic factors that make them an excellent candidate for use in the clinical setting to provide cell-based restoration of hyaline-like cartilage. Although allogeneic in nature, these cells stimulate little or no host immune response and can be stored for long periods while maintaining viability. We present a technique of cartilage repair in the knee using Wharton's jelly–derived MSCs embedded onto scaffolding and implanted in a minimally invasive fashion using dry arthroscopy.

Human umbilical cord Wharton jelly cells promote extra-pancreatic insulin formation and repair of renal damage in STZ-induced diabetic mice

BackgroundWe evaluated the therapeutic effect and fate of high doses of human umbilical cord Wharton jelly cells (hUCWJCs) after IP administration to streptozotocin (STZ)-induced diabetic mice.MethodsType 1 diabetes (T1D) was induced in Kunming mice via IP injection of STZ. hUCWJCs were labeled with 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI). Diabetic animals with sustained hyperglycemia for at least 2 weeks were administered 1 × 107 Dil-hUCWJCs via intraperitoneal injection. Insulin, glucagon and PDX-1 were detected by immunofluorescence with confocal microscopy. Serum mouse and human C-peptide was assayed in blood collected via intracardiac puncture. Specific β-cell differentiation markers and human DNA were assessed using qPCR performed with 200 ng of target DNA.ResultshUCWJCs migrated to the STZ-damaged organs and contributed to lower blood glucose levels in 30% of the treated mice. Confocal microscopy revealed the presence of resident insulin-positive cells in the liver and kidneys. hUCWJC-treated mice with restored hyperglycemia also showed increased serum mouse C-peptide levels. The qPCR results, particularly in the liver, revealed that after transplantation hUCWJCs upregulated genes of endocrine precursors but failed to express endocrine stage markers. Mice with restored hyperglycemia had reduced urinary volume and lacked glomerular hypertrophy, exhibiting a morphology resembling that of normal glomeruli. Moreover, we also verified that one of the possible mechanisms by which hUCWJCs exert immunosuppressive effects is through down-regulation of the cell surface receptor HLA-1.ConclusionsWe confirmed the potential of IP administration of hUCWJCs and the capability of these cells to migrate to damaged tissues and promote insulin secretion from non-pancreatic local cells and to improve renal damage. These findings confer unique therapeutic properties to hUCWJCs, suggesting a promising future in the treatment of diabetes mellitus.

Runt-Related Transcription Factor 2 Induction During Differentiation of Wharton's Jelly Mesenchymal Stem Cells to Osteoblasts Is Regulated by Jumonji AT-Rich Interactive Domain 1B Histone Demethylase.

Novel bone regeneration approaches aim to obtain immature osteoblasts from somatic stem cells. Umbilical cord Wharton's jelly mesenchymal stem cells (WJ-MSCs) are an ideal source for cell therapy. Hence, the study of mechanisms involved in WJ-MSC osteoblastic differentiation is crucial to exploit their developmental capacity. Here, we have assessed epigenetic control of the Runt-related transcription factor 2 (RUNX2) osteogenic master regulator gene in WJ-MSC. We present evidence indicating that modulation of RUNX2 expression through preventing Jumonji AT-rich interactive domain 1B (JARID1B) histone demethylase activity is relevant to enhance WJ-MSC osteoblastic potential. Hence, JARID1B loss of function in WJ-MSC results in increased RUNX2/p57 expression. Our data highlight JARID1B activity as a novel target to modulate WJ-MSC osteoblastic differentiation with potential applications in bone tissue engineering. Stem Cells 2017;35:2430-2441.

Differentiation of mesenchymal stem cells to germ-like cells under induction of Sertoli cell-conditioned medium and retinoic acid.

The aim of this research was to find a way to differentiate germ cells from umbilical cord Wharton's jelly mesenchymal stem cells (MSCs) to support in vitro spermatogenesis. A small piece of Wharton's jelly was cultured in high-glucose Dulbecco's modified Eagle's medium in present of 10% foetal calf serum. After the fourth passage, the cells were isolated and cultured in Sertoli cell-conditioned medium under induction of two different doses of retinoic acid (10-5 , 10-6 m). The differentiation of MSC to germ-like cells was evaluated by expression of Oct4, Nanog, Plzf, Stra8 and Prm1 genes during different days of culture through qPCR. The results showed that there were downregulation of Oct4 and Nanog and upregulation of pre-meiotic germ cell marker (stra8) and haploid cell marker (Prm1) when MSCs are differentiated over time. The expression of Bax gene (an apoptotic marker) was significantly observed in high dosage of retinoic acid (RA). As a result, RA has positive effects on proliferation and differentiation of MSCs, but its effects are related to dosage. The success of this method can introduce umbilical cord MSC as a source of germ cells for treatment of infertility in future.

Induction of Immunogenic Cell Death in Lymphoma Cells by Wharton's Jelly Mesenchymal Stem Cell Conditioned Medium.

Strategies that induce immunogenic cell death (ICD) or downregulate CD47 or PD-L1 expression have resulted in successful therapeutic options for tumor eradication. Several groups have reported the tumoricidal effects of human umbilical cord Wharton's jelly stem cells (hWJSCs) or its conditioned medium (hWJSC-CM) on certain cancers but the mechanisms have not been elucidated. Since hWJSCs possess immunomodulatory properties, we investigated whether one of the tumoricidal mechanisms was via ICD. We first concentrated hWJSC-CM into a 3kDa concentrate and then exposed various concentrations of this concentrate to human lymphoma cells to find out which concentration had the greatest tumoricidal effect. We observed that a 500µg/ml concentration of the concentrate had the greatest inhibitory effect. Thereafter, lymphoma cells were exposed to 500µg/ml of the hWJSC-CM-3kDa concentrate and then subjected to analysis for morphology, viability, apoptosis, mitochondrial and endoplasmic reticulum stress, danger associated molecular patterns (DAMP), extracellular HMGB1, CD47 and PD-L1 markers and dendritic cell phenotype. Extensive nuclear chromatin and mitochondrial changes with significantly decreased cell viability and increased apoptosis were observed in the treated lymphoma cells compared to controls. There were also significant increases in the release of DAMPs, extracellular HMGB1 and dendritic cell activation and maturation, with concomitant decreases in CD47 and PD-L1 expression in the treated cells compared to controls. In other ongoing studies we observed increased expression of specific tumor-suppressor molecules (miRNA-146a and miRNA-126, MCP-1, IL-6, IL-8 and IL-12) in hWJSC-CM suggesting that one or more of these molecules may be the modulators of the ICD.

Correlation of CDC42 Activity with Cell Proliferation and Palmitate-Mediated Cell Death in Human Umbilical Cord Wharton's Jelly Derived Mesenchymal Stromal Cells.

RHO GTPases regulate cell migration, cell-cycle progression, and cell survival in response to extracellular stimuli. However, the regulatory effects of RHO GTPases in mesenchymal stromal cells (MSCs) are unclear. Herein, we show that CDC42 acts as an essential factor in regulating cell proliferation and also takes part in lipotoxic effects of palmitate in human umbilical cord Wharton's jelly derived MSCs (hWJ-MSCs). Cultured human bone marrow, adipose tissue, and hWJ-MSC derived cells had varying pro-inflammatory cytokine secretion levels and cell death rates when treated by palmitate. Strikingly, the proliferation rate of these types of MSCs correlated with their sensitivity to palmitate. A glutathione-S-transferase pull-down assay demonstrated that hWJ-MSCs had the highest activation of CDC42, which was increased by palmitate treatment in a time-dependent manner. We demonstrated that palmitate-induced synthesis of pro-inflammatory cytokines and cell death was attenuated by shRNA against CDC42. In CDC42 depleted hWJ-MSCs, population-doubling levels were notably decreased, and phosphorylation of ERK1/2 and p38 MAPK was reduced. Our data therefore suggest a mechanistic role for CDC42 activity in hWJ-MSC proliferation and identified CDC42 activity as a promising pharmacological target for ameliorating lipotoxic cell dysfunction and death.

Вплив низьких температур на біологічно активні сполуки вартонієвого студня пуповини людини

Вплив низьких температур на біологічно активні сполуки вартонієвого студня пуповини людини