目的:建立大鼠肝细胞无血清原代培养体系。方法:以Wistar大鼠做肝细胞供者,采用胶原酶肝脏原位灌流消化法分离肝细胞,分别通过有血清和无血清方法进行原代培养;以台盼蓝染色法测细胞活力,倒置显微镜观察肝细胞形态变化,流式细胞仪测定原代培养大鼠肝细胞增殖指数。结果:在大鼠肝脏有血清原代培养体系和无血清原代培养体系两种方法中,大鼠肝细胞存活率均>90%,生长良好,经24 h和48 h培养后,经检测大鼠肝细胞增殖指数无统计学差异(P>0.05)。结论:成功建立大鼠肝细胞无血清原代培养方法,为今后利用此方法进行细胞信号传导等相关研究提供有力的实验手段。 Objective: To establish a serum-free primary culture system of rat hepatocytes. Methods: Hepatocytes were donated by Wistar rats. Hepatocytes were isolated by collagenase hepatic in situ perfusion digestion method, primary cultured by serum-free and serum-free method. Cell viability was measured by trypan blue staining. The morphological changes of hepatocytes were observed. The proliferation index of primary cultured rat hepatocytes was determined by flow cytometry. RESULTS: In the two methods of primary serum-free and serum-free primary culture in rat liver, the survival rate of rat hepatocytes was> 90%, and grew well. After being cultured for 24 h and 48 h, Rat hepatocyte proliferation index no significant difference (P> 0.05). CONCLUSION: The successful establishment of rat serum-free primary culture of rat hepatocytes provides a powerful experimental method for future studies on cell signal transduction using this method.