人胚肝细胞分离培养研究  

Studies on Isolation and Primary Culture of Human Fetal Hepatocytes

作　　者：黄柏英[1] 余剑英[2] 祝和成[1] 

机构地区：[1]中南大学湘雅医学院细胞生物学研究室 [2]湘雅医院核医学科,湖南长沙410078

出　　处：《湘南学院学报（自然科学版）》2005年第1期10-11,14,共3页

摘　　要：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。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.

关 键 词：人胚 分离培养 聚乙烯吡咯烷酮(pvp) 胶原酶消化法 胰蛋白酶 原代肝细胞 细胞存活率 肝细胞移植 优化培养 生长能力 消化方法 细胞生长 体外培养 人工肝 

分 类 号：Q813[生物学—生物工程] S858.28[农业科学—临床兽医学]