微载体黏附培养SD大鼠肝细胞的生物学特性

Study of biological characteristics of microcarrier-attached SD rat hepatocytes

目的 :探讨微载体黏附培养SD大鼠肝细胞的生物学特性 .方法 :对胶原酶消化法分离获取的SD大鼠肝细胞行微载体黏附培养 ,在倒置显微镜及扫描电镜下观察肝细胞形态变化 ,采用CL 80 0全自动生化分析仪检测不同时间培养上清中白蛋白、乳酸脱氢酶 (LDH)和尿素的含量 .结果 :微载体黏附培养肝细胞的正常形态及白蛋白、尿素合成功能可维持 1wk以上 ,LDH漏出量、白蛋白及尿素水平 1wk内呈波动性变化 ,在培养第 3日白蛋白及尿素水平最高、LDH漏出量最低 .结论 :微载体培养可提供高活性、高密度生长的肝细胞 ,可望为肝细胞移植、生物型人工肝提供较理想的细胞材料 ;微载体培养肝细胞在培养第 3日功能最佳 ,可能为应用于肝细胞移植。

AIM: To explore the biological characteristics of microcarrier attached SD rat hepatocytes. METHODS: The SD rat hepatocytes obtained by collagenase digestion were cultured on microcarrier. The morpholgical changes were examined by reverse microscopy and scanning electron microscopy, and the levels of albumin, LDH, urea were detected by CL 800 auto biochemical analysis. RESULTS: The morphological characteristics and abilities of albumin and urea synthesis remained unchanged for more than 1 week. The LDH leakage, albumin synthesis and urea level were found to be fluctuated within one week,and there was a less LDH leakage and higher albumin and urea levels on the third day. CONCLUSION: Microcarrier culture can provide hepatocytes with high density and viability. Microcarrier attached hepatocytes canbe a good cell material for hepatocyte transplantation and bioartificial liver support system (BLSS). As the microcarrier attached rat hepatocytes have the best function on the third day, the third day may be the optimal time of applying microcarrier culture hepatocytes for hepatocyte transplantation and BLSS.