金黄地鼠原代肝细胞的分离纯化与培养

目的 建立一种简单、高效的金黄地鼠肝细胞分离、纯化及培养的方法.方法 实验采用两步胶原酶原位灌流法分离地鼠原代肝细胞,percoll离心法纯化细胞,台盼蓝染色检测成活率,对所分离培养的原代肝细胞进行形态学鉴定,Western Blot检测细胞功能.结果 经过分离纯化的金黄地鼠肝细胞,细胞成活率为（93.7％±2.1）％,细胞形态呈现不规则多边形,细胞多为双核和多核细胞.细胞在腺苷处理后,AMP激活的蛋白激酶（AMPK）磷酸化水平明显升高.结论 成功建立了金黄地鼠肝细胞分离纯化及培养的方法.

Isolation, purification and culture of Hamster primary hepatocytes

Objective To establish an easy and efficient method of the isolation, purification and culture of Hamster primary hepatocytes. Methods Hamster Primary Hepatocytes were isolated using two-step collagenase perfusion in situ and purified by centrifugation with Percoll. The yields and viabilities were measured by standard trypan blue exclusion. Hepatocytes were identified using morphologic observation. Cell function was evaluated by Western blot method. Results Under the optical microscope, the obtained hepatocytes exhibited as irregular polygon with two or more circular nucleuses in each cell. The average viability of hepatocytes was 93.7% ± 2.1%. After incubated with adenosine, intracellular AMPK phosphorylation was significantly increased. Conclusion The method for isolating, purifying and culturing Hamster primary hepatocytes was successfully established.