Transdifferentiation of Human Umbilical Cord Stromal Cells to Neurogenic-Like Cells

Abstract

Objective: To investigate the differentiation of human umbilical cord stromal cells into neurogenic cells. Methods: (1) Human umbilical cord stromal cells (HUMSCs) were pre-induced with recombinant human fibroblast growth factor-2 (hbFGF-2, Peprotech, UK) and N² (1:100, Invitrogen, USA), immunocytochemistry and Western blot to determine the expression of nestin and fibronectin before and after pre-induction. Western blot to identify Nestin and fibronectin expression before and after pre-induction. (2) The differentiation of pre-induced and un-induced HUMSCs into neurogenic cells was performed, and the value-added rates of both were measured by MTT at different time points, and the morphological differences between the differentiated cells and the differentiated cells were verified at the gene and protein levels. Results:(1) 94.4 ± 2.3% and 6.5 ± 1.2% HUMSCs expressed ectodermal marker-fibronectin and stem cell marker-nestin, respectively, and 62.3 ± 5.2% piHUMSCs showed nestin immunopositive staining after adding FGF2 and N² for pre-induction; 51.3 ± 5.9% of cells showed fibronectin immunopositive staining. piHUMSCs showed significantly higher nestin positivity than HUMSCs (P<0.05), whereas HUMSCs expressing fibronectin showed significantly higher positivity than piHUMSCs (P<0.05). Western blot results were consistent with cellular immunohistochemistry results. The proliferation rate of HUMSCs and piHUMSCs by MTT assay suggested that after 48 h of culture, both HUMSCs and piHUMSCs entered the logarithmic cell growth phase, and the cell expansion rate of HUMSCs was significantly higher than that of piHUMSCs after 72 h and 96 h of culture.(2) The results of RT-PCR experiments indicated that differentiated human umbilical cord neural stem cells expressed The expression of the corresponding neural stem cell marker genes was statistically different (p<0.05) than that of undifferentiated umbilical cord stromal cells and terminally differentiated neural cells after induction of fractionation. Conclusion: Umbilical cord-derived stromal cells pre-induced with recombinant human fibroblast growth factor-2 (hbFGF-2, Peprotech, UK) and N² (1:100, Invitrogen, USA) were better able to differentiate into neural stem cells with stable expression of cell morphological changes, proteins and genes.