Small molecules cocktail enhances endothelial differentiation of human gingiva mesenchymal stem cells

Abstract

Objectives: Endothelial cells-based blood vessel engineering could be one solution to treat ischemic diseases. However, the scarcity of source and limited expansion rate restrict the translational application of primary autologous endothelial cells (ECs). In the present study, we aim to use small molecule cocktail-based protocol to induced human gingiva mesenchymal stem cells (GMSCs) into endothelial-like cells, providing an alternative source of ECs for the treatment of ischemic diseases. Methods: Passage 3-5 GMSCs were cultured in EGM2-MV with a set of small molecule compounds and growth factors to generate endothelial-like cells (GMSC-ECs). Endothelial specific gene and protein (CD31, VEGFR2, VE-CADHERIN, vWF) expression was detected using RT-PCR, western blotting, flow cytometry, and immunofluorescence. GMSC-ECs were further compared with primary human umbilical vein endothelial cells (HUVECs) in terms of their functions in vitro, such as tubular formation assay and LDL-uptake evaluation. Results: GMSC-ECs cells exhibited strong expression of EC-specific markers. In vitro tubular formation assay demonstrated that the GMSC-ECs derived vascular structure was similar to those of HUVECs. GMSC-ECs could also incorporate ac-LDL in vitro. Conclusions: Endothelial differentiation of GMSCs was significantly promoted by small molecules cocktail. The GMSC-ECs could be promising candidate cells for novel therapy of ischemic vascular diseases.