大鼠原代肝脏细胞的分离培养及其膜受体鉴定

Isolation and Culture of Rat Primary Hepatocytes,and Identification of its Membrane Receptor

本研究采用改良的两步胶原酶灌流法分离培养大鼠肝脏细胞,并对其进行了形态学观察、功能检测及表面受体的鉴定,以探索其短期培养的最佳功能状态。结果表明,平均每只大鼠可获取2.12×108个肝脏细胞,平均活率达94.23%;光镜下观察发现肝脏细胞呈多角形并成片生长;对肝脏细胞的LDH、Alb和尿素进行连续检测,结果发现细胞离体培养第3天功能较好;Mab263-PE单抗鉴定结果显示,肝脏细胞生长激素受体功能健全尚未受损,为细胞信号转导等相关试验奠定了基础。

This research adopted improved collagenase perfusion isolation to isolate and culture rat hepatocytes, and had carried on the observation of morphology,identification of function and membrane receptor to explore the optimal state of its short-term cultures.The results showed that 2.12×108 hepatocytes per rat liver were obtained, and the average motility rate of cells was 94.23% .Hepatocytes were polygonal and grew into flakinesst under light microscope.We found that hepatocytes culture in vitro were better on the third day by the continual detection of LDH,Alb and urea.Function of growth hormone receptor on liver membrane had not been damaged by the identification of Mab263-PE,which laid the foundation for the cell signal transduction and relevant test.