大量分离乳猪肝细胞的方法  

Large scale isolation of suckling pig hepatocytes

作　　者：王英杰[1] 刘国栋[1] 刘俊[1] 李梦东[1] 

机构地区：[1]第三军医大学西南医院传染病专科中心,重庆市400038

出　　处：《世界华人消化杂志》1999年第8期661-662,共2页World Chinese Journal of Digestology

基　　金：国家"九五"攻关项目

摘　　要：目的　探讨大量分离乳猪肝细胞的方法．方法　用体外灌流装置，ＥＤＴＡ 和胶原酶两步灌流法消化分离乳猪肝细胞，综合方法观察判断肝细胞产量、活力、纯度及增殖能力．结果　分离肝细胞的总产量达到１０９／ 肝，细胞活力９４ ％ ～９８ ％ ，纯度大于９６ ％ ． 分离乳猪肝细胞胞膜完整、细胞器丰富，培养肝细胞有活跃的增殖能力，培养上清中乳酸脱氢酶浓度仅有轻度升高，２４ ｈ 平均浓度１７－８ Ｕ／Ｌ±５－４ Ｕ／Ｌ．AIM To study the method for large scale isolation of suckling pig hepatocytes. METHODS Liver cells of suckling pig were harvested by two step perfusion with ethulenediaminetetraacetate and collagenase by using an extracorporeal perfusion apparatus. The cell yield, viability, purity, and proliferation were observed and detected respectively. RESULTS The total yield of isolating hepatocytes reached to 10 9 in each pig liver with the viability of 94% to 98% and purity of more than 96%. Isolated hepatocytes demonstrated by transmission electron microscopy showing integrity of plasma membranes and intracellular organelles. Cultured hepatocytes isolated by the method proliferated significantly. Lactate dehydrogenase in the media showed a slight increase. CONCLUSION This method allows isolation of large numbers of suckling pig hepatocytes with higher viability.

关 键 词：肝细胞 细胞分离 生物人工肝 

分 类 号：R318.14[医药卫生—生物医学工程]