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目的 :建立稳定的猪肝细胞的分离与原代培养体系 ,满足生物人工肝体外支持系统对肝细胞的需求。方法 :应用改良 Seglen法进行肝脏原位胶原酶灌流分离猪肝细胞 ,进行平面培养 ,动态观察细胞生长过程中的形态结构变化。结果 :平均每只猪可获得 2× 1 0 1 0个肝细胞 ,细胞活率达到 92 .5 %。在平面培养过程中维持了正常肝细胞的形态。结论 :原位胶原酶灌流法是获得大量高活率的猪肝细胞的首选方法 ,猪肝细胞原代分离培养是目前解决组合型生物人工肝脏 (HBL SS)应用中细胞来源问题的主要途径
OBJECTIVE: To establish a stable isolation and primary culture system of porcine hepatocytes to meet the demand of hepatocytes by the bioartificial extrahepatic support system. Methods: Segmental liver cells were isolated by hepatic collagenase in situ hybridization with modified Seglen method. The morphological changes of the cells were observed dynamically by plane culture. Results: On average, 2 × 10 000 hepatocytes were obtained per pig, and the viability of the cells reached 92.5%. In the plane culture process to maintain the normal morphology of liver cells. CONCLUSION: In situ collagenase perfusion method is the preferred method to obtain a large number of high viable porcine hepatocytes. The primary isolation and culture of porcine hepatocytes is the main approach to solve the problem of cell source in combined bioartificial liver (HBL SS)