人胚肝细胞分离培养研究

目的　探讨体外优化培养人胚肝细胞的方法。方法　采用胰蛋白酶和聚乙烯吡咯烷酮(pvp) ,胶原酶,胶原酶和pvp消化法分离人胚肝细胞,并比较3种方法对分离人胚肝细胞的效果。结果　运用这三种方法均能成功地培养出原代肝细胞,并能够传代;但胶原酶和pvp消化法分离细胞效果最好,细胞贴壁生长能力强。三种消化方法其细胞存活率从高到低依次为胶原酶和pvp消化法、胶原酶消化法、胰蛋白酶和pvp消化法。结论　人胚肝细胞的分离首选胶原酶和pvp消化法,使用该法在体外培养的人胚肝细胞生长良好,可用于肝细胞移植和生物人工肝等方面的研究。

Studies on Isolation and Primary Culture of Human Fetal Hepatocytes

Objective To study the Culture methods of human fetal hepatocytes. Methods Trypsin and pvp,collagenase,collagenase and pvp digestive methods were used to isolate human fetal hepatocytes (HFH) and the efficiency of three kinds of digestive methods for isolating HFH were compared.Results The primary culture of HFH has been successfully established by the use of three methods and the cells were passaged. The results sho wed that satisfactory isolation of HFH and good attachment were achieved by using the collagenase and pvp digestive method, The cells viability of three digestive methods from high to low were collagenase and pvp digestive method, collagenase digestive method, Trypsin and pvp digestive method.Conclusions Collagenase and pvp digestive method shoud first be selected in isolating HFH, the HFH cultured in this experiment survived well and can serve the further studies of liver cell transplantation and bioartificial liver support system.