体外培养条件下肝细胞功能研究

Functional study of hepatocytes cultured in sandwich configuration

目的 探讨一种新的细胞培养模型以使体外培养的肝细胞更好地恢复肝功能。方法利用三明治构型培养肝细胞 ,观察体外肝细胞蛋白合成、生物转化及其mRNA表达并与传统单层胶原培养肝细胞相比较。结果 三明治构型培养肝细胞保持稳定的蛋白分泌功能并持续增长 ,且有良好的生物转化功能 ,而传统单层胶原培养肝细胞蛋白分泌功能微弱 ,7d后基本消失 ,肝细胞生物转化功能差。原位杂交显示三明治构型培养较单层胶原培养肝细胞具有更强的白蛋白、P4 50 bmRNA表达。结论 三明治构型培养肝细胞较传统单层胶原培养肝细胞更类似体内肝细胞 ,可体外重建细胞极性。

Objective To develop a new culture system in which hepatocytes can restore in-vivo like function. Methods Sandwich configuration was used for the culture of hepatocytes, and protein synthesis as well as P 450-b ,albumin mRNA expression by in situ hybridization was studied. ResultsStable and gradually increasing protein synthesis and high level of P 450-b , albumin mRNA expression were observed. In contrast, protein secretion of hepatocytes cultured in conventional single collagen culture system was low and decreased until approximately zero seven days after, and mRNA expression of P 450-b ,albumin was low. Conclusions Sandwich configuration in which cell polarity of hepatocytes is reestablished like in vivo situation.