The Relative Study Of The Process That VEGF-C Synergizes HGF Promote The Differentiation From Mesenchymal Stem Cells Into Lymphatic Endothelial Cells

Objective:To explore the optimal concentration of Hepatocyte Growth Factor (HGF) and the mechanism of marrow stem cells (BMSCs) differentiated into lymphatic endothelial cells (LECs) by HGF synergizeing with VEGF-C. Methods:Isolating and culturing the BMSCs. Using flow cytometry (FCM) detect the relative Surface antigen of rat BMSCs and identificated the ability of multipotent differentiation. The 3th generation of rat BMSCs were used in the experiment divided into 6 synergizeing groups including 50ng/ml VEGF-C synergizeing with 10,20,40,60,80, 100ng/ml HGF each, and set 50ng/ml VEGF-C inducing group,50ng/ml HGF induing group, blank control inducing group. After 10d inducing, Lymphatic endothelial hyaluronan receptor 1(LYVE-1) and Prospero homeobox protein 1(Prox1), the cell makers of LECs were detected using RT-qPCR. Then HGF inducing group, VEGF-C alone inducing group and the blank control group were set in detecting the cell makers of LECs afterlOd inducing by using Western Blot and RT-qPCR. Lastly, blank control group, VEGF-C inducing group and HGF synergizeing with VEGF-C inducing group were set in detecting integrinα9 after 10d inducing by using Western Blot and RT-qPCR. Results:Purification of rat bone marrow mesenchymal stem cells were successfully isolated and cultured. The of LYVE-1、Prox1 were the highest expression (PProx1=0.000, PLYVE-1=0.000) in the inducing group of 80ng/ml HGF synergizeing with VEGF-C. Meanwhile, The HGF inducing group were not detected LECs markers, and comparing HGF synergizeing with VEGF-C inducing group with VEGF-C inducing group,the former has the higher expression of integrinα9 (Pwestern=0.001, PRT-qPCR=0.000). Conclusion:HGF can synergize with VEGF-C better enable the rat BMSCs differentiate into LECs when the concentration of HGF is 80ng/ml, but it could not induce rat BMSCs differentiate into LECs alone at this concentration. HGF can up-regulate integrinα9 expression indirectly in the process of differentiation which point out that HGF may play a role in the up-regulation of integrinα9 expression indirectly to promote the differentiation.