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目的研究用微载体培养人肝细胞及肝非实质细胞的方法．方法采用体外简易两步灌流和差速离心法获取胎肝细胞和肝非实质细胞，在综合限定条件下进行微载体ｃｙｔｏｄｅｘ３粘附培养，并对培养肝细胞的形态及合成葡萄糖和白蛋白的功能进行动态测定．结果分离肝细胞及肝非实质细胞在接种时即呈明显的聚集倾向，将其与微载体混合振荡孵育２０ｍｉｎ后，二者极易相粘附．在被覆聚羟乙基异丁烯酸的培养瓶内，使用激素限定条件培养液培养约４８ｈ，典型的多细胞聚集球形体得以形成．该形态特征以及白蛋白、葡萄糖合成能力可保持１月结论使用微载体培养人肝细胞和肝非实质细胞具有多种优点，有广泛的应用价值．
Objective To study the method of culturing human hepatocytes and non-parenchymal cells with microcarriers. Methods Fetal hepatocytes and non-parenchymal cells were obtained by simple two-step in vitro perfusion and differential centrifugation. The cells were cultured with microcode cytodex3 under the comprehensive conditions. The morphology of cultured hepatocytes and the function of synthesizing glucose and albumin Dynamic measurement. Results The isolation of hepatocytes and non-parenchymal cells showed obvious aggregation tendency at the time of inoculation. After 20 min incubation with microcarriers, the two were easily adhered. In polyhydroxyethylmethacrylate-coated flasks, culture was performed for approximately 48 hours using hormone-conditioned medium, and typical multicellular aggregated spheroids were formed. The morphological characteristics and albumin, glucose synthesis ability can be maintained in January Conclusion The use of microcarrier cultured human hepatocytes and non-parenchymal cells has many advantages, has a wide range of applications.