中文摘要：

目的 建立一种简单的能获得高纯度小鼠原代肝细胞的分离与培养方法。方法 将经典的两步灌流法简化为经下腔静脉逆向脉冲式灌注,利用低速离心法对肝细胞分离液进行纯化;利用锥虫蓝染色法鉴定细胞活率;培养瓶预先用鼠尾胶原铺备,利用倒置显微镜观察细胞形态。结果 此法得到了足够数量的肝细胞,细胞活率大于90%;细胞可稳定贴壁1周。结论 成功建立了一种简单的分离与培养小鼠原代肝细胞的方法。

英文摘要：

Objective To create a simple method of isolating and culturing high-purity mouse primary hepatic cells. Methods The classic two-step perfusion technique was simplified to intermittent retrograde perfusion from the inferior vena cava. The cell suspension was purified by a low speed centrifugation. The motility rate was assessed by trypan blue staining. The culture flask was pre-paving by mouse tail collagen. The morphous of the hepatocytes was observed by an invert microscope. Results Adequate hepatocytes were obtained. The motility rate of the hepatocytes was higher than 90%. The viability of the cells could last for one week. Conclusion A simple method of isolation and primary culture of mouse hepatocytes has been successfully created.