Study Of The ADSCs Promotion Of Remnant Liver Regeneration After Excessive Hepatectomy

[Objective] To explore the rat ADSCs acquisition method in vitro,biological characteristics,the capacity of differentiation in vitro and differentiate into liver cell in the model of excessive hepatectomy,in order to explore the ADSCs promotion of liver regeneration.[Methods]â‘ We fetch inguinal fat of male SD rats to primary culture, observe cell growth and cell morphology, depict cell growth curve.(2)Take the third generation cell and flow cytometry identified this cells and determine cell purity.â‘¢reparation of conditioned induced medium in vitro,use the third generation cell to prepared cell climbing film,add induced medium culture cell,after 14,21 d immunohistochemistry detected cell Alb and CK18.â‘£Prepared the model of excessive hepatectomy,through the branches of the portal vein injected the male ADSCs,after 1,7,14,21 d check liver function(AST,ALT,TB,A1b,TP),routine pathological examination of liver tissue and immunohistochemistry detected Y chromosome sry gene, respectively Alb,CK18 double positive cells after 14,21d.[Results]â‘ The cell covered the bottom about 10 days after primary culture,the cell were fibroblast-like or vortex-like, with the more passage times, the cell morphology more single, cell growth curve like"S". The third generation cells had the strongest proliferation.â‘¡By flow cytometry to detect cell surface markers, CD44 expression rate about 98.5%, CD90 expression rate about 92.6%, CD49d expression rate about 90.5%, CD29 expression rate about 73.1%, CD45 expression rate about 0.3%, CD34 expression rate about 0.6%. Through the combination between them surface markers confirm this cell is ADSCs, and has high purity.â‘¢Cell climbing film began morphological changes from long fusiform into edge shape or oval through 5 days induce and some cell appeared dual-core, this is unique to liver cells. Alb and CK18 expression positive on 14,21days, and with the induction time increased gradually extended.â‘£The experimental group than the control group and the negative control group ALT, AST decreased significantly after cell transplantation 21 days,it has Statistics meaning(P<0.05). The experimental group and the control group, the negative control group has no significant difference at each time by measured TB(P >0.05), each group TP, Alb numerically first increased and then decreased with time over, the experimental group than the control group and the negative control group TP, Alb increased significantly after 21 days and the difference was significant(P<0.05). Immunohistochemistry detected liver tissue of the experimental group Alb, CK18 and the Y chromosome sry gene in double-stained cells showed positive expression of 14 days, the number of positive cells increased 21 days.[Conclusion]â‘ DSCs can be got by a special cultured method from the adipose tissue, and cultured in vitro can be passaged for many times, a lot of amplification, while maintaining its biological characteristics.Their has a very strong proliferation, and the third generation has strongest proliferation.â‘¡onfirm these cell is ADSCs by flow cytometry detected a variety of cell surface antigens and the combination of many different surface antigens,and these cell has high purity.â‘¢ADSCs can differentiate into hepatocyte-like cells in a suitable culture medium,and express the liver cell antigen,it has the function of liver cells.â‘£DSCs can differentiate into hepatocyte-like cells in the model of excess hepatectomy, differentiation cell expressed the hepatocyte-specific antigen and has the function of liver cells, it contribute to the recovery of liver function after liver injury and promote liver regeneration after excessive hepatectomy.