Apoptosis Related Human Wharton’s Jelly-Derived Stem Cells Differentiation into Osteoblasts, Chondrocytes, Adipocytes and Neural-like Cells—Complete Transcriptomic Assays

Abstract

Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) exhibit multilineage differentiation potential, adhere to plastic, and express a specific set of surface markers—CD105, CD73, CD90. Although there are relatively well-established differentiation protocols for WJ-MSCs, the exact molecular mechanisms involved in their in vitro long-term culture and differentiation remain to be elucidated. In this study, the cells were isolated from Wharton’s jelly of umbilical cords obtained from healthy full-term deliveries, cultivated in vitro, and differentiated towards osteogenic, chondrogenic, adipogenic and neurogenic lineages. RNA samples were isolated after the differentiation regimen and analyzed using an RNA sequencing (RNAseq) assay, which led to the identification of differentially expressed genes belonging to apoptosis-related ontological groups. ZBTB16 and FOXO1 were upregulated in all differentiated groups as compared to controls, while TGFA was downregulated in all groups. In addition, several possible novel marker genes associated with the differentiation of WJ-MSCs were identified (e.g., SEPTIN4, ITPR1, CNR1, BEX2, CD14, EDNRB). The results of this study provide an insight into the molecular mechanisms involved in the long-term culture in vitro and four-lineage differentiation of WJ-MSCs, which is crucial to utilize WJ-MSCs in regenerative medicine.

Expression Profile of New Marker Genes Involved in Differentiation of Human Wharton’s Jelly-Derived Mesenchymal Stem Cells into Chondrocytes, Osteoblasts, Adipocytes and Neural-like Cells

“…After the third passage, the WJ-MSCs were subjected to osteogenic, chondrogenic, adipogenic and neurogenic differentiation. The results of the differentiation were confirmed with specific staining and presented in previous articles [19]. Briefly, in adipoinduced WJ-MSCs stained with Oil Red O, the red lipid droplets were observed, which were not present in the control sample.…”

“…Several researchers suggested that the in vitro differentiation of MSCs might decrease the sensitivity of these cells to apoptosis after the transplantation [ 24 , 25 ], while the others obtained the opposite outcomes [ 26 ]. Our previous results obtained after performing RNA-seq on differentiated WJ-MSCs with four lineages indicated that differentiation of WJ-MSCs prior to the transplantation would not be more beneficial than the utilization of undifferentiated cells, considering the expression of apoptosis-related genes in these groups [ 19 ]. Conflicting results emphasize the importance of investigation of the molecular processes occurring during the differentiation of MSCs, not only those engaged in the apoptotic process, to decide whether the differentiation conducted prior to the transplantation of cells would be beneficial.…”