Wharton's Jelly-derived Mesenchymal Cells Research Articles

Differential gene expression of Wharton's jelly-derived mesenchymal cells mediated by graphene oxide in basal and osteo-induced media.

Graphene oxide (GO) is widespread in scaffold engineering owing to its extraordinary properties such as multiple oxygen functional groups, high hydrophilicity ability and biocompatibility. It is known to promote differentiation in mesenchymal stem cells, but concomitant comparison of its modulation on the expression profiles of Wharton's jelly (WJ)-MSC surface markers, lineage differentiation, and epigenetic regulatory genes in basal and induced condition are still lacking. Unraveling the fundamental mechanisms is essential for the effective utilization of WJ-MSCs incorporated with GO in therapy. This study aims to explore the unique gene expression profiles and epigenetic characteristics of WJ-MSCs influenced by GO. The characterized GO-coated coverslip served as a substrate for culturing WJ-MSCs. In addition to investigating the impact of GO on cell proliferation and differentiation, we conducted a gene expression study using PCR array, while epigenetic control was assessed through bisulfite sequencing and Western blot analysis. Our findings indicate that the presence of GO maintained the proliferation and survival of WJ-MSCs. In the absence of induction, GO led to minor lipid and glycosaminoglycan deposition in WJ-MSCs. This was evidenced by the sustained expression of pluripotency and lineage-specific genes, demethylation at the OCT4 promoter, and a decrease in H3K9 methylation. In osteo-induced condition, the occurrence of osteogenesis appeared to be guided by BMP/TGF and ERK pathway activation, accompanied by the upregulation of osteogenic-related genes and downregulation of DNMT3b. GO in osteo-induced condition create a favorable microenvironment that promotes the osteogenesis of WJ-MSCs by influencing genetic and epigenetic controls. This helps in advancing our knowledge on the use of GO as priming platform and WJ-MSCs an alternate source for bone repair and regeneration.

Mesenchymal Cells Retain the Specificity of Embryonal Origin During Osteogenic Differentiation.

Mesenchymal stem cells (MSCs) are widely used in therapy, but the differences between MSCs of various origins and their ability to undergo osteogenic differentiation and produce extracellular matrix are not fully understood. To address this, we conducted a comparative analysis of mesenchymal cell primary cultures from 6 human sources, including osteoblast-like cells from the adult femur, adipose-derived stem cells, Wharton's jelly-derived mesenchymal cells, gingival fibroblasts, dental pulp stem cells, and periodontal ligament stem cells. We analyzed these cells' secretome, proteome, and transcriptome under standard and osteogenic cultivation conditions. Despite the overall similarity in osteogenic differentiation, the cells maintain their embryonic specificity after isolation and differentiation in vitro. Furthermore, we propose classifying mesenchymal cells into 3 groups: dental stem cells of neural crest origin, mesenchymal stem cells, and fetal stem cells. Specifically, fetal stem cells have the most promising secretome for various applications, while mesenchymal stem cells have a specialized secretome optimal for extracellular matrix production. Nevertheless, mesenchymal cells from all sources secreted core bone extracellular matrix-associated proteins. In conclusion, our study illuminates the distinctive characteristics of mesenchymal stem cells from various sources, providing insights into their potential applications in regenerative medicine and enhancing our understanding of the inherent diversity of mesenchymal cells in vivo.

Study of the Cellular Senescence Process in Human Umbilical Cord Wharton's Jelly-derived Mesenchymal Stem Cells

Introduction: Embryonic stem cells are candidates for the treatment of regenerative medicine, but their use is faced with limitations due to ethical issues. The human umbilical cord-derived mesenchymal stem cells (MSCs) are appropriate options because the cells have no ethical difficulties and have self-renewal. Senescence is known as a gradual functional loss that heterogeneously occurs in multiple organ systems. Objective: The objective of this study was to investigate the cellular senescence process in human umbilical cord Wharton's jelly-derived mesenchymal cells. Materials and Methods: Umbilical cord was obtained from healthy newborns at the General Hospital of Tabriz. Under sterile conditions, Wharton's jelly was removed from the blood vessels and minced into small pieces of about 0.5 mm. These were cultured in Dulbecco's Modified Eagle's Medium (MSC medium). Real-time polymerase chain reaction for p16INK4a and senescence-associated β-galactosidase (SA-β-gal) staining was performed to investigate the cellular senescence process. Results: The results showed the different expressions in the different passages, but it was significantly increased from the fifth passage compared to the first passage. SA-β-gal staining also showed increased colour intensity in the fifth passage compared to the first passage. Conclusion: SA-β-gal is not a specific marker for senescence, while p16INK4a is a specific marker. Further studies are required for the investigation of the senescence mechanism, such as the evaluation of genes involved in the senescence.

A novel hybrid polymer of PCL/fish gelatin nanofibrous scaffold improves proliferation and differentiation of Wharton's jelly-derived mesenchymal cells into islet-like cells.

Using a different source of stem cells to compensate for the lost beta cells is a promising way to cure diabetic patients. Besides, the best efficiency of insulin-producing cells (IPCs) will appear when we culture them in an environment similar to inside the body. Hence, three-dimensional (3D) culture ameliorates the differentiation of diverse kinds of stem cells into IPCs compared to those differentiated in two-dimensional (2D) culture. In this study, we aim to create an ideal differentiation environment by using PCL/Fish gelatin nanofibrous scaffolds to differentiate Wharton's jelly-derived mesenchymal cells (WJ-MSCs) to IPCs and compare them with a 2D cultured group. The evaluation of cellular, molecular, and functional properties of differentiated cells on the 3D and 2D cultures was investigated by several assays such as electron microscopy, quantitative PCR, immunochemistry, western blotting, and ELISA. The in vitro studies showed that WJ-MSCs differentiated in the 3D culture have strong properties of IPCs such as islet-like cells. The expression of pancreatic-specific genes at both RNA and protein levels showed higher differentiation efficacy of 3D culture. Besides, the results of the ELISA tests demonstrate that in both groups the differentiated cells are functional and secreted C-peptide and insulin in glucose stimulation, but the secretion of C-peptide and insulin in the 3D culture group was higher than those cultured in 2D groups. Our findings showed the use of PCL/Fish gelatin nanofibrous scaffolds with optimized differentiation protocols can promote the differentiation of IPCs from WJ-MSCs compared to the 2D culture group.

Effects of polarized and non-polarized red-light irradiation on proliferation of human Wharton's jelly-derived mesenchymal cells

Light emitting diode (LED) irradiation has recently been introduced as an encouraging strategy for promotion of cell proliferation. Human umbilical cord Wharton's jelly-derived mesenchymal (hUCM) cells are among the most available mesenchymal cells with a promising application in regenerative medicine. The aim of the present study was to examine the effect of polarized (PL) and non-polarized (NPL) red-light emitted by LED on various proliferation properties of hUCM cells. Cell proliferation was assessed 48 h after irradiation of hUCM cells by different energy densities. Cell density increased to a significant level both in PL and NPL irradiation at 0.954 J/cm2 following WST-1 assay. Staining of irradiated and non-irradiated cells with Hoechst after 3 and 6 days revealed an increased proliferation rate in irradiated cells, but the non-irradiated cells proliferated more than irradiated cells at day 9 of cultivation. Similar results were obtained in trypan blue assay. Scratch repair test for 18 h with an interval of 6 h did not reveal a significant difference between irradiated and non-irradiated cells. In addition, CFU-F assay in PL irradiated cells was higher than control when 500 cells/plate was cultivated. Totally, this study revealed that hUCM cells could be induced to achieve higher number of cells by PL and NPL red-light irradiation after 48 h.

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.