剑尾鱼肝细胞原代培养  

Primary Culture of Swordtail Fish(Xiphophorus helleri) Hepatocytes

作　　者：梁岳[1] 马广智[1] 方展强[1] 

机构地区：[1]华南师范大学生命科学学院,广州510631

出　　处：《中国比较医学杂志》2006年第3期185-187,F0002,共4页Chinese Journal of Comparative Medicine

基　　金：广东省自然科学基金项目(04010378);广东省科技计划项目(2004B40101015)

摘　　要：目的探讨剑尾鱼肝细胞原代培养的方法.方法分别用机械分散法、酶消化法对肝细胞进行分离,并比较在William＇s E（Williams＇Medium E）、DMEM、DMEM/F12无血清培养基中肝细胞的生长效果.结果 （1）组织块培养法（机械分散法）的肝细胞在William＇s E、DMEM、DMEM/F12无血清培养基中,温度25℃～28℃,pH 7.3左右时都可以短期正常生长,而生长效果好依次是William＇s E、DMEM、DMEM/F12无血清培养基.（2）酶消化法分离的肝细胞的产量为（6.4±1.7）×106 cell/g肝组织,即时存活率为（85±6）%.24 h贴壁率不足20%.结论酶消化法培养肝细胞产量、即时存活率及贴壁率偏低.组织块培养法肝细胞生长良好,而不需要苛刻的条件,可以满足毒理学实验要求.培养肝细胞首选William＇s E培养基.Objective To study the primary culture methods of hepatocytes of swordtail fish （Xiphophorus helleri ）. Methods Hepatocytes were isolated by digestion with pancreatin and mechanical separation. The growth effect of hepatocytes cultured in William＇s E （William＇ s Medium E）, DMEM and DMEM/F12 （Serum-free medium） was compared. Results （1） With mechanical separation method, hepatoeytes could normally grow in all the mediums of William＇s E（ William＇s Medium E）, DMEM, DMEM/F12 （Serum-free medium） at the temperature 25℃ ± 28℃, pH7.3 in a short-term. The best growth of hepatocytes was in William＇ s E, followed by culture in DMEM, and least in DMEM/F12. （2） With digestion with pancreatin, the average yield of hepatocytes was （6.4 ± 1.7） × 10^6 cell/g liver tissue, and the average viability was （85 ± 6） % . The rate of cell adherence was not more than 20% at 24 h. Conclusion The results indicated that the average yield, the average viability and the rate of cell adherence of hepatocytes isolated by digestion method with pancreatin was lower. However, hepatocytes culture obtained by mechanical separation method could grow well and deos not need rigorous conditions, and can well satisfy the requirement of toxicological experiments. It also indicated that the William＇ s Medium E is a preferred medium for the primary culture of Xiphophorus helleri hepatocytes.

关 键 词：鳉形目 肝细胞 细胞 培养 剑尾鱼 原代培养 

分 类 号：R394[医药卫生—医学遗传学]