Effect of mesenchymal stem cells on differentiation potential of blood endothelial progenitor cells

GE Quanhu; WU Xiangwei; CHENG Wenzhe; WAN Longfei; WANG Xiaoyi; ZHAO Bowen

doi:10.3969/j.issn.1002-266X.2017.28.004

Abstract：Objective To investigate the effect of mesenchymal stem cells (MSCs) on the differentiation of endothelial progenitor cells (EPCs) into endothelial cells (ECs).Methods MSCs and EPCs were isolated, cultured, and amplified from murine bone marrow in vitro.MSCs and EPCs were cultured in Transwell co-culture system.The experiment was totally divided into three groups, EPCs cultured group (control group), co-culture group, and vascular endothelial growth factor group (VEGF group).After being cultured for 48 h, EPCs were collected and the expression of the specific markers of eNOS and KDR was measured by RT-PCR;the expression of VEGF in the culture medium of each group was detected by ELISA.MSC conditioned medium (MSCCM) was collected.EPCs which were at passage 3 were cultured in the medium of LG-DMEM, MSCCM, MSCCM+IgG, MSCCM+VEGF neutralizing antibody (Anti-VEGF), respectively, which were taken as the following four groups, LG-DMEM group, MSCCM group, MSCCM+IgG group, and MSCCM+Anti-VEGF group.After being cultured for 48 h, EPCs were collected and the expression of eNOS and KDR was measured by RT-PCR.Results The mRNA expression of the specific markers was significantly higher in the co-culture group and VEGF group than that in the control group (P<0.05 or <0.01).The expression of VEGF in the co-culture group and VEGF group was significantly increased as compared with that of the control group (P<0.01).The mRNA expression of the specific markers in the MSCCM group and MSCCM+IgG group was significantly higher than that of the LG-DMEM group and MSCCM+Anti-VEGF group (P<0.01).Conclusion MSCs can promote the differentiation of EPCs into ECs through paracrine factors such as VEGF.