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目的 建立乳猪肝细胞与非实质细胞大量聚集培养技术。探讨获取大量高活性乳猪肝细胞的培养方法。方法 采用磁力悬浮培养技术和装置进行聚集培养。观察聚集体的形态 ,受损伤程度和白蛋白分泌功能 ,氨转化功能。结果 一次培养肝细胞数可达 2× 10 10 ～ 5× 10 10 个 ,聚集体肝细胞结构完整 ,相互之间形成胆管样结构等肝组织特异结构 ,保持良好生物活性。结论 聚集培养技术将细胞接种浓度提高 5倍 ,一次可制备足够数量的肝细胞 ,较好地满足构建杂合型人工肝时对生物材料的要求
Objective To establish a technique for accumulating large numbers of hepatocytes and non-parenchymal cells in suckling pigs. To explore a large number of highly active suckling pig hepatocytes culture method. Methods Magnetic suspension culture technology and apparatus were used for aggregation culture. Aggregate morphology was observed, the degree of damage and albumin secretion, ammonia conversion function. Results The number of primary cultured hepatocytes reached 2 × 10 10 ~ 5 × 10 10. The aggregated hepatocytes were characterized by their complete structure and the specific structures of the liver such as bile duct-like structures and their biological activities. Conclusion Aggregation culture technology can increase the concentration of cells inoculation by a factor of 5, and can prepare a sufficient number of hepatocytes at one time to better meet the requirements of biomaterials in the construction of heterozygotic artificial liver