肝细胞生长因子诱导小鼠胚胎干细胞分化的实验研究

Study on differentiation of murine embryonic stem cells into hepatocyte-functional cells induced by hepatocyte growth factor (HGF)

目的探讨肝细胞生长因子(HGF)诱导小鼠胚胎干细胞(ESC)体外定向分化为肝细胞的能力.方法对ESC体外悬浮培养5～7天形成的拟胚体,观察ESC自主分化、HGF诱导分化的情况.观察和检测经诱导4周的细胞的HE染色,糖原染色,尿素氮及甘油三酯合成,心肌MHC、白蛋白、AFP、CK18的表达,以及吲哚氰绿(ICG)染色情况.结果 ESC自主分化难以控制.HGF可促进ESC向内胚层进一步分化,但更可能诱导其向心肌分化.表达白蛋白、AFP、CK18、ICG和FDA染色阳性.结论 HGF可诱导ESC分化出肝细胞,但HGF的单一作用能力有限,提示肝细胞的诱导分化可能需要多种因素共同作用.

Objective To investigate the functional role of murine hepatocyte growth factor （HGF） in inducing in vitro the mouse embryonic stem cells （ESC） to differentiate unidirectionally into hepatocyte. Methods Both spontaneous and HGF induced differentiations of the primordial cell elusters formed by suspended culture in vitro of ESC were daily observed with microscope for 5-7 days. After 4 weeks of culture, the cells were stained with HE and glycogen staining, their morphology were observed, The synthesis of urea nitrogen and triglycerides in the culture medium, the expression of myocardial MHC, albumin, AFP and CK18, and the indocyanine green （ICG） staining were also detected. Results It was hard to control the stmntaneous differentiation of E~. HGF could promote the differentiation of ESC into endoderm, and more likely into myocardium. HGF could also induce the expression of albumin, AFP and CK18, and positive staining of ICG and FDA. Conelusions HGF may induce the differentiation of ESC into hepatocyte, but the functional role is limited. It implies that a comprehensive effort of multiple factors might be needed in inducing the hepatocyte differentiation.