The Differentiation Of Bone Mesenchymal Stem Cells To Hepatocyte In Hepatectomined Mouse

Objective: To investigate the methods of isolating bone mesenchymal stem cells (BMSCs) from BALB/C mouse and whether BMSCs could different into hepatocytes and participate in regeneration in partly hepatectomined mouse.Methods: BMSCs were isolated from the bone marrow of BALB/C male donor mouse by adherence selecting in DMEM media with low glucose and 20% fetal bovine serum. The primary and the 5th passage cells (P5) were analyzed with flow cytometry. 48 female mice were studied and divded to three group averagely-control group, model group and transplant group. Model group and transplant group were hepatectomined, and transplant group immediately accepted BMSCs transplant.Liver tissues were gathered and manufactured tissue microarray. Simultaneous detection were performed on regenerated liver for both the Y-chromosome by fluorescence in situ hybridization and the albumin or cytokeratin 18 (CK18) by fluorescence immunoassays. The fluorescence was objected by general fluorescence microscope and confocal microscope. Image analyzed by laser sharp 2000 or photoshop 7.5. The liver pathology, serum biochemistry, avoirdupois and the weight of liver were analyzed to confirm whether the transplant of BMSCs can help the liver regeneration inhepatectomined mouse.Results: Cells stick the wall after 80 minutes and stretched as spindle after 48 hours. 98.7 % of the primary bone cells didn't express CD44 and CD29. No the primary bone cell expressed CD44 and CD29 simultaneously. 97.4% P5 adherence cells expressed CD29. 80.8% P5 adherence cells expressed CD44 and CD29 simultaneously and 16.6% P5 adherence cells only expressed CD29.At 5th day after partly hepatectomine, avoirdupois of model mice and transplant mice decreased compared with that in control mice. At 14th day after partly hepatectomine, there was no difference among the three groups. At 14th day after partly hepatectomine, the weight of liver and the index of liver weight in model mice decreased compared with that in control group, whereas there was no difference between transplanted mice and control mice.At 5th day after partly hepatectomine, there were no difference of total bile (TBil), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) among transplanted mice, model mice and control mice. Serum total protein (TP) and albumin (ALB) decreased in transplanted mice and model mice compared with that in control mice, whileas globulin (GLOB) increased in transplanted mice and model mice compared with that in control mice. At 14th day after partly hepatectomine, there were no difference of TBil, ALT, TP and ALB among transplanted mice, model mice and control mice. GLOB decreased in transplanted mice and model mice compared with that in control mice. AST in transplant mice increased dramatically compared with that in model mice and control mice. The vessel endothelium cell which had shuttle shape and acidophilic plasma detected in transplant mice.The cells which showed both Y chromosome and albumin (or CK18)positive can be detected in both rejected liver lobe and other lobe at 5th day and 14th day after hepatectomine and transplant. These cells located in hepatic plate. Most of these cells exist separate, but some cells are close or form a node. Some of these cells have two Y chromosome in karyon.Conclusion:1 CD4/ CD29+ cells and CD44" CD29+ cells can be isolated by the condition of 48 hours adherence, 20% fetal bovine serum and DMEM with low glucose from bone marrow.2 BMSCs could proliferate and migrate in liver lobe, and different into hepatocytes, and participate in liver regeneration.3 The security of BMSCs transplant need more study.