成年鸡原代肝细胞两种分离方法的比较  

Comparison of two culture methods of primary chicken hepatocytes

作　　者：刘静[1] 刘传敦[1] 丁雪莹[1] 唐兆新[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医杂志》2012年第6期34-38,共5页Chinese Journal of Veterinary Medicine

摘　　要：为获得数量多及活力好的原代肝细胞,本试验分别采用原位二步法与半原位法分离鸡的肝细胞。应用MTT法比较了两种分离方法的细胞活力,应用电镜观察了分离细胞的超微结构,并分别在体外培养的第1、3、5天和第7天检测肝细胞上清液的AST、ALT、ALB和GGT,以鉴定鸡原代肝细胞的生化功能。结果显示,采用半原位法分离的鸡肝细胞与原位二步法相比,细胞数量多、活力高、结构完整,可在体外持续培养7d并稳定地保持肝细胞的形态与功能。To obtain a large number of highly viable primary hepatocytes from adult chicken,the two-steps perfusion method and the semi-situ perfusion method were applied to isolate hepatocytes,respectively.The cell viability of hepatocytes was determined by the MTT method,the morphology was observed using electric microscope,and the metabolism was determined by AST,ALT,ALB,and GGT assay.Conclusively,the semi-situ perfusion method was better in getting larger amount and higher viability（P〈0.05） of hepatocytes in comparison with the two-steps perfusion method,and the cells can be cultured for at least 7 days with normal morphology and function.

关 键 词：鸡肝细胞 半原位法 原位二步法 原代培养 

分 类 号：S852.2[农业科学—基础兽医学]