Shi Li-jun, Li Shuang-xing, Sun Bo, Wang Jing-hua, Li Hu-lun, Jin Liang-hong
Department of Gastroenterology, First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Zhonghua Gan Zang Bing Za Zhi. 2007 Sep;15(9):681-4.
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.
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 alpha-actin (alpha-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.
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 alpha-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).
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.
研究骨髓间充质干细胞（BMSCs）的分化以及BMSCs对体外肝硬化贮脂细胞（CFSC）和肝细胞增殖的影响。
从大鼠骨髓和肝脏中分离并收获BMSCs和肝细胞。通过Transwell小室建立共培养体系，两个小室由半透膜分隔。将用PKH26标记的BMSCs与肝细胞/CFSC在共培养体系中以及细胞-细胞直接接触培养体系中培养。采用荧光免疫细胞化学检测抗白蛋白和抗平滑肌α-肌动蛋白（α-SMA）抗体。单独培养的BMSCs和肝细胞/CFSC作为对照。检测共培养体系中肝细胞的增殖水平。用BMSCs的条件培养基培养CFSC，并在显微镜下测量其数量。
两个培养体系中的肝细胞培养72小时后均观察到白蛋白表达，但细胞-细胞直接接触培养体系中的白蛋白水平更高（P<0.01）。与对照组相比，共培养体系中的肝细胞数量更多（P<0.01）。在任一培养体系中均未观察到CFSC中α-SMA的表达。BMSCs的条件培养基抑制CFSC的增殖。共培养时间越长，CFSC生长抑制越显著（P<0.01）。
BMSCs可被肝细胞形成的局部微环境诱导分化为肝细胞。BMSCs可能促进肝细胞增殖并抑制CFSC增殖。
