摘要
目的探讨微环境对骨髓基质干细胞（BMSCs）分化的影响以及BMSCs对肝纤维化细胞（CFSC）和肝细胞增殖的作用。方法分离、培养大鼠原代BMSCs和肝细胞,用PKH26荧光标记BMSCs,将标记后的BMSCs与肝细胞/CFSC进行接触和非接触共培养,用抗白蛋白抗体/抗α、平滑肌抗体对BMSCs进行检测;BMSCs条件培养液与CFSC共培养,倒置显微镜下计数CFSC的细胞数量变化。结果BMSCs与肝细胞共培养72h,白蛋白染色均出现阳性,且接触共培养的BMSCs白蛋白染色阳性率高于非接触共培养（P〈0.01）。肝细胞与BMSCs非接触共培养48h后,肝细胞的数量明显多于对照组（P〈0.01）。BMSCs与CVSC共培养体系中BMSCs的形态无明显变化,且αSMA染色未出现阳性。BMSCs条件培养液能抑制CFSC的增殖,作用时间越长,抑制作用越明显（P〈0.01）。结论肝细胞形成的局部微环境能诱导BMSCs向肝样细胞分化,BMSCs能够促进肝细胞生长并对CFSC的增殖有明显的抑制作用。
Objective To investigate the differentiation of bone marrow mesenchymal stem cells （BMSCs） and the effects of BMSCs on the proliferation of cirrhotic fat-storing cells （CFSC） and hepatocytes in vitro. Methods BMSCs and hepatocytes were isolated and harvested from the bone marrow and livers of rats. A co-culture system was set up by transwell inserts in which the two chambers were separated by a semipermeable membrane. BMSCs labeled with PKH26 were cultured with hepatocytes/CFSC in the co- culture system and also in a cell-cell direct contact culture system. Anti-albumin and anti-smooth muscle α - actin （ α -SMA） antibodies were tested by using fluorescence immunocytochemistry. BMSCs and hepatocytes/CFSC cultured alone served as controls. The proliferation level of hepatocytes in the co-culture system was measured. CFSC were cultured with the conditional medium of BMSCs, and their quantities were measured microscopically. Results Expression of albumin was observed in the hepatocytes of the two culture systems after they were cultured for 72 h but the albumin levels were higher in the cell-cell direct contact culture system （P 〈 0.01 ）. As compared to the controls, the number of hepatocytes was larger in the co-culture system （P 〈 0.01）. No expression of α -SMA in CFSC was observed in either culture system. The proliferation of CFSC was inhibited by the conditional medium of BMSCs. The longer the time of the co-culturing the more significant was the CFSC growth suppression （P 〈 0.01 ）. Conclusions BMSCs can be induced into hepatocytes by a local micro-environment formed by hepatocytes. BMSCs may promote proliferation of hepatocytes and inhibit proliferation of CFSC.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2007年第9期681-684,共4页
Chinese Journal of Hepatology
基金
黑龙江省博士后基金（LRD-2005-35175）
关键词
干细胞
肝细胞
共同培养
肝纤维化细胞
Stem cells
Hepatocyte
Co-culture
Cirrhotic fat-storing cells
