壳聚糖球形多孔微载体支持下培养的肝细胞功能及其代谢活性检测

Function and metabolic activity of hepatocytes cultured by spherical porous chitosan microcarriers

背景:在肝细胞移植及生物型人工肝研究中,肝细胞培养仍然是其关键,如何方便地获取足够数量、活性较好、功能良好的肝细胞已经成为其首要问题。微载体培养技术作为一项体外高密度细胞培养技术,近年来已在肝细胞的体外培养中得到应用。目的:对壳聚糖球形多孔微载体培养的人肝细胞L-02进行定时的细胞功能和代谢活性检测。方法:对以自制的壳聚糖球形多孔微载体样本为支架培养的人肝细胞L-02进行定时的细胞功能检测(包括测定谷草转氨酶,谷丙转氨酶和乳酸脱氢酶质量浓度)和代谢活性检测(包括检测培养基中白蛋白、尿素、葡萄糖含量),其中实验组为壳聚糖球形多孔微载体支持下培养人肝细胞L-02,对照组为无壳聚糖球形多孔微载体支持下培养人肝细胞L-02,检测时间点为人肝细胞L-02培养的第1,2,3,4,5天。结果与结论:实验组和对照组测定的谷草转氨酶、谷丙转氨酶和乳酸脱氢酶活性在前3d持续下降,第3天降到最低,而从第4天开始重新反弹上升;但白蛋白、尿素和葡萄糖水平在前3d持续上升,第3天升到最高值,而从第4天开始逐渐下降,实验组每天测定上述各项水平始终明显高于对照组(P<0.05),提示实验组中的人肝细胞L-02代谢活性较对照组增强。

BACKGROUND： Hepatocyte culture is the key to the hepatocyte transplantation and BAL study. It has turned into the first problem how to get adequate numbers, better activity and well-functioning hepatocyte. In the recently years, the microcarrier as a technique for high density cell culture has been applied in the in vitro culture of hepatocytes. OBJECTIVE： To detect the function and metabolic activity of human hepatocytes L-02 cultured by spherical porous chitosan microcarriers at each time. METHODS： The function, including aspartate aminotransferase （AST）, alanine aminotransferase （ALT） and lactate dehydrogenase （LDH） levels, and metabolic activity, including albumin （ALB）, urea, glucose contents, of human hepatocytes L-02 cultured by spherical porous chitosan microcarriers were detected at 1, 2, 3, 4, 5 days of culture. RESULTS AND CONCLUSION： In the two groups, the levels of AST, ALT and LDH decreased at 1-3 days, and increased at 4 days; the contents of ALB, urea and glucose increased at 1-3 days, and decreased at 4 days. The levels of all the indexes in the experimental group were obviously higher than those in the control group （P 0.05）. It indicated that the metabolic activity of human hepatocytes L-02 in the experimental group was higher than that in the control group.