肝细胞体外培养技术的研究与进展

目前肝细胞体外培养技术主要有以下几种：单层培养法、胶原凝胶三明治培养法、微载体黏附培养法、微囊包裹培养法、球形聚集培养法、中空纤维培养法。胶原凝胶三明治培养法制备简单，培养效果明确，但是所培养的细胞数量有限，难于大规模工业化培养；中空纤维培养法能为细胞生长提供了三维空间，增加了肝细胞与培养体系的接触面积，有利于物质代谢与交换，但存在管上细胞分布欠均匀、细胞团块易堵塞半透膜孔隙影响物质交换等弊端；球形聚集培养法不需构建载体，但球体大小不易控制，易造成球体中心肝细胞坏死；微囊包裹培养能提供一个较好的免疫阻隔环境，能在免疫阻隔要求较高的环境下使用。微载体黏附培养法能够在单位体积内培养最多数量的肝细胞，能对肝细胞的黏附、生长和分泌提供较好环境，目前微载体的直径和孔径还未达到最理想的培养要求，单位体积肝细胞的培养数量还有很大的提升空间。同时培养液配方与共同培养技术的不断改良，必将使体外培养的肝细胞数量与功能达到肝细胞移植所需。

Research and progress of in vitro culture method of hepatocytes

At present, the main methods of culture in vitro of hepatocytes are as followings: flat-plate culture, collagen sandwich culture, microcarrier culture, microcapsule culture, spheroid culture and hollow fiber culture. The preparation of sandwich culture system is simple and the method has clear effect of cultivation. However, by the limited amount of cultured cells, it is difficult to cultivation in large-scale industrialization. The hollow fiber culture can provide a three-dimensional environment so that the hepatocytes cultured inside can get a greater contact surface with the cultivation system and enhance metabolism and exchange of material. But hepatocytes cultured on the surface of the hollow fibers are less evenly distributed and easily clump the semipermeable membrane to impact on the exchange of material. The spheroid culture does not need to construct carriers, but it is hard to control the diameter of the cell balls which result in the necrosis in the centre of the ball. The microcapsule culture can provide a better immunization barrier environment which can be used in higher demand of immune barrier. Microcarrier culture can culture the maximum number of hepatocytes in unit volume and provide a better environment for hepatocytes to adhesion, growth and secretion. Nowadays, the diameter and the aperture of the microcarrier have not yet reached an ideal level. The quantity of hepatocytes in unit volume still has a great potential of increasing. With the improvement of medium ingredient and coculture technology, the culture in vitro of hepatocytes will meet the requirements of hepatocyte transplantation in sufficient amount and ideal performance.