肝细胞生长因子诱导华通胶干细胞向肝细胞样细胞分化的体外实验

摘要

Objective To observe the capability of hepatocyte growth factor in inducing Wharton's jelly -derived mesenchymal stem cells ( WJMSCs) into hepatocyte -like cells. Methods Wharton's jelly - derived mesenchymal stem cells were isolated from human umbilical cord. The phenotype of mesenchymal stem cells and telomerase activity were identified. Four groups were set up to investigate the capability of hepatocyte growth factor in inducing Wharton's jelly - derived mesenchymal stem cells into hepatocyte - like cells j Group A: WJMSCs + HGF(20 ng/ml) ; Group B: WJMSCs + HGF(40 ng/ml) ; Group C: HL -7702 cell line and Group D: WJMSCs| , and evaluate preliminarily by detecting the relative gene expression quantity of AFP and ALB. Results The phenotype of Wharton's jelly - derived mesenchymal stem cells was positive in CD73 , CD90, CD105 and HLA - ABC, but it was negative in CD45 and HLA - DR. Telomerase activity of WJMSCs showed no significant difference compared with that of HeLa cells. The mRNA of AFP increased at the 7th day in group A and B, while it was lower in group A than that of group B ( P < 0. 05). They both showed much lower than that of group C (P < 0. 05) , while there was no significant difference at 21 st and 42nd day in group C (P >0. 05 ) . The mRNA of ALB increased at the 7th day in group A and B ( P > 0. 05) , but they were both lower than that of group C ( P < 0. 05). At 21 st day, ALB in group A and B showed no significant difference compared with that of group C (P > 0. 05) ; and it was higher in group A and B than that of group C at the 42nd day (P <0. 05). Conclusion Hepatocyte growth factor could induce Wharton's jelly - derived mesenchymal stem cells into hepatocyte - like cells, and this effect is related to the concentration of hepatocyte growth factor in the culture medium.%目的 观察外源性肝细胞生长因子对华通胶干细胞向肝细胞样细胞分化的诱导作用.方法 取人脐带华通胶干细胞,进行细胞表型分析和端粒酶活性评价；通过检测华通胶干细胞中甲胎蛋白(AFP)和白蛋白(Alb)两种基因的相对含量,对两种浓度肝细胞生长因子诱导能力进行初步评估.分组:A组:华通胶干细胞+肝细胞生长因子(HGF)(20 ng/ml)；B组:华通胶干细胞+HGF(40 ng/ml)；C组:HL-7702人肝细胞株；D组:华通胶干细胞.结果 华通胶干细胞表达间充质细胞表型:CD73、CD90、CD105与HLA-ABC阳性；不表达造血干细胞表型:CD45、HLA-DR阴性；华通胶干细胞端粒酶表达呈阳性.AFP基因相对含量在第7d表达增高,仍未达到阳性对照组水平(P＜0.05)；高剂量组合量高于低剂量组(P＜0.05)；在21、42 d两组相对含量与阳性对照组差异无统计学意义；Alb基因相对含量在第7d表达增高,高剂量与低剂量组差异无统计学意义,低于阳性对照组(P＜0.05)；在21 d两组相对含量与阳性对照组差异无统计学意义；在42 d,两组相对含量高于阳性对照组(P＜0.05).结论 肝细胞生长因子可以诱导华通胶干细胞向肝细胞样细胞进行分化,该诱导作用与肝细胞生长因子剂量有关.