摘要目的开发从屠宰器官进行高活性猪肝细胞规模分离的优化程序.方法采用部分肝叶逆行灌注、机械及组酶肝消化并经密度梯度离心获取高产量、高纯度肝实质性细胞.结果部分肝叶经消化可获取1.39×109肝细胞的平均产量(9.9×106肝细胞/克组织)及92.5%的平均细胞存活率.经组合性dispase/胶原酶的消化处理肝细胞的收获量及存活率明显增加.对细胞灌流及分离液进行氧化处理能阻止泡状细胞的发生.分离的猪肝细胞经原代培养其活性及机能保持着与鼠肝细胞一致的水平,且随时间的推移而减退.结论通过我们改良的方法可收获高产量成年猪肝细胞,并保持良好的活性与分化机能.因此,成年猪肝细胞被证实是人工肝脏辅助支持系统中生物组的一个很有价值的肝细胞来源.

abstractsObjective　To develop procedures for the successful harvesting of large quantities of viable and functional pig liver cells from abattoir organs.Methods　The procedure included partial liver lobe retrograde perfusion and mechanical/enzymatic digestion of the liver tissue, followed by separation of the hepatocytes, based on size and density, from contaminating cell types.Results　Digestion of the partial liver lobe resulted in an average yield of 1.39×109 cells (9.9×106 cells/g liver) with an average viability of 92.5%. The yield and viability of cells were improved by dispase/collagenase resultant digestion. The emergence of blebby cells was blocked by supplying oxygen to the cell isolation buffers. Isolated hepatocytes seeded onto polystyrene surfaces remained viable and functional at a level comparable to that of rat hepatocytes, although their function decreased over time.Conclusions　Adult pig hepatocytes can be harvested with high yields and retain viability and differentiated function using this method. Abattoir pig livers can be an excellent source of hepatocytes for use as the biological component of artificial liver assist devices.