猪肝细胞的分离与原代培养

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2×10¹⁰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（HBLSS）应用中细胞来源问题的主要途径。

Isolation and primary cultivation of porcine hepatocytes

Objective To establish the isolation and primary cultivation system of porcine hepatocytes in accordance with the requirement of hybrid bioartificial liver support system (HBLSS) of cell in vitro .Methods Porcine hepatocytes were isolated with digestive method of collagenase perfusion with the improved Seglen method. Morphological changes of the hepatocytes were observed.Results The average yield of hepatocytes was 2×10 10 cells per pig and the average viability was 92 5%.Conclusion The digestive method of collagenase perfusion is the basic approach for obtaining a mass of hepatocytes with high viability. Porcine hepatocytes are the prime cells source for HBLSS.