一种SD大鼠肝细胞原代培养的改良方法

目的 建立一种改良的大鼠肝细胞原代培养方法.方法取6到7周龄SD 大鼠（约200g）,3%戊巴比妥钠麻醉,采用Seglen两步胶原酶灌流法分离肝细胞,Percoll密度梯度离心纯化肝细胞,种植在预先用gelatin处理的培养皿内.结果 分离所得肝细胞成活率达95%以上.24h后用相差显微镜观察细胞贴壁良好,细胞形态规则;48h后细胞开始伸展.结论 使用改良的方法获得的细胞成活率高,贴壁牵,适合常见的药理毒理实验.

An improved method for primary culturehepatocytesof SD rat

Objective To establish an improved method for rat hepatocyte primary culture. Methods SD rat （6 - 7 weeks old, about 200g） was anaesthetized with 3% pentobarbital sodium, seglen two - step collagenase perfusion method and percoU density gradient centrifugation were used to isolate and purify hepatocytes. The harvested liver ceils were planted in the Petri dishes pre - processed with gelatin. Results Survival rate of the isolated hepatocyteswas above 95%. Cells were found to be adhered well to the plate under phase contrast microscope after 24h, the morphology of cells present well. Cells started stretch 48hafter plantation. Conclusions We got high survival rate hepatocytes by using this new method, it could be applied in pharatacology and toxicology experiment.