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目的:初步探讨不同浓度的肝细胞生长因子(hepatocyte growth factor,HGF)和成纤维细胞生长因子(fibroblast growth factor,FGF)对大鼠肝干细胞增殖的调控作用。方法:先将大鼠肝胎细胞分别种植A、B培养板上进行肝细胞的体外培养。A组:培养液中含有HGF,浓度分为别1,5,10,20,40,80及100ng/mL。B组:培养液中含有FGF,浓度分别为1,5,10,20,40,80及100ng/mL。对照组不加细胞因子。由A、B两组剂量效应获得HGF和FGF的最佳浓度。再将细胞接种于C组培养板进行培养,培养液中含有最佳浓度HGF联合上述各浓度FGF,并用四甲基偶氮唑盐比色法(mononuclear cell direct cytotoxicity assay,MTT)分别测两组的光密度值(optical delnsity,OD),检测出HGF与FGF的最佳浓度组合。结果:HGF和FGF浓度与肝干细胞增殖效应呈剂量依赖性(dose-dependent)。当HGF浓度为20ng/mL,FGF浓度为10ng/mL时,为最佳浓度组合。结论:HGF和FGF均能对肝干细胞的增殖起促进作用,并且两者有一定协同作用。
Objective: To investigate the effects of different concentrations of hepatocyte growth factor (HGF) and fibroblast growth factor (FGF) on the proliferation of hepatic stem cells in rats. Methods: First, rat hepatic fetal cells were planted on A and B plates for hepatocyte culture in vitro. Group A: The culture medium contains HGF at the concentrations of 1, 5, 10, 20, 40, 80 and 100 ng / mL, respectively. Group B: FGF was contained in the culture medium at concentrations of 1, 5, 10, 20, 40, 80 and 100 ng / mL, respectively. Control group without cytokines. The optimal concentrations of HGF and FGF were obtained from the dose effects of A and B groups. Then the cells were inoculated in group C culture plates for culture. The culture medium contained the best concentration of HGF in combination with each of the above concentrations of FGF, and the two groups were respectively measured by MTT (mononuclear cell direct cytotoxicity assay) The optical density (OD) of HGF and FGF was detected. Results: The concentrations of HGF and FGF were dose-dependent with the proliferation of hepatic stem cells. When HGF concentration of 20ng / mL, FGF concentration of 10ng / mL, the best concentration combination. Conclusion: Both HGF and FGF can promote the proliferation of hepatic stem cells, and they both have some synergistic effects.