VEGF-C combined with HGF promotes the differentiation of mesenchymal stem cells into lymphatic endothelial cells

Abstract:

Objective：To explore the optimal concentration of hepatocyte growth factor（HGF） and the mechanism of marrow stem cells（BMSCs） differentiate into lymphatic endothelial cells（LECs） by HGF synergizeing with vascular endothelial growth factor C（VEGF-C）. Methods：BMSCs were isolated and cultured. Flow cytometry（FCM） was used to detect the relative surface antigen of rat BMSCs and the ability of multipotent differentiation was identified. The 3th generation of rat BMSCs were used in the experiment and were di－vided into 6 synergizeing groups including 50 ng/ml VEGF-C synergizeing with 10，20，40，60，80，100 ng/ml HGF and 50 ng/ml VEGF-C inducing group，50 ng/ml HGF induing group，blank control inducing group were established. After 10 d inducing，lymphatic endothelial hyaluronan receptor 1（LYVE-1） and prospero home－obox protein 1（Prox1），the cell makers of LECs were detected using real-time PCR. Prox1 and LYVE-1 in blank control group，VEGF-C inducing group and HGF inducing group after10d in－ducing were detected by using Western blot and real-time PCR. Expression of integrinα9 in blank control group，VEGF-C in－ducing group and HGF synergizeing with VEGF-C group was detected by Western blot and real-time PCR. Results：Purifica－tion of rat bone marrow mesenchymal stem cells was successfully isolated and cultured. The expressions of LYVE-1 and Prox1 were the highest（PProx1=0.000，PLYVE-1=0.000） in 80 ng/ml HGF synergizeing with VEGF-C group. Meanwhile，LECs markers were not detect－ed in HGF inducing group.Higher expression of integrinα9 was observed in HGF synergizeing with VEGF-C group than in VEGF-C inducing group（PWestern blot=0.001，Preal-time PCR=0.000）. Conclusion：HGF synergizeing with VEGF-C can better enable rat BMSCs differen－tiate into LECs when the concentration of HGF is 80 ng/ml，but HGF alone could not induce rat BMSCs differentiate into LECs at this concentration. HGF can up-regulate integrinα9 expression indirectly in the process of differentiation，indicating that HGF may promote the differentiation through up-regulating integrinα9 expression indirectly.