| Liver is the most important energy metabolism, biosynthesis and material conversion organ of human being.Serious illness of liver always needs OLT ,but the donors are limited,and the operation is difficult and expensive.They make the OLT difficult to widely use. Hepatocyte transplantation have several virtues:a donor can provid cells for several receptors,the operation of HTx is easy,the cells are low immunogenicity and can be saved by freezer,the transplantation can be done by several times,it is little harm for receptors if the operation failed.HTx is limited by the lack of human liver cells and serious immunological rejection of different species liver cells. Hepatic stem cell have the ability of differentiation into several directions.HSC can differentiate into hepatocytes and other liver cells.It is a new source of hepatocytes by more and more studies of HSC. Mesenchymal stem cells have the ability of differentiation into several directions.MSC is named extrahepatic HSC.MSC can differentiate into oval cells,hepatocytes and biliary hepatocytes.The properts of MSC make the use of patien's MSC to self-transplantation opportunity.It can solve the main problems of the HTx,the lack of human liver cells and serious immunological rejection after transplantation ,at the same time.Objective :1. We isolate human embryonic MSC and colture it in vitro,and fumble about the way of isolation and colture.2.We establish the animal model of hepatocytes anti- regeneration and liver injure.3.We transplante human embryonic MSC into the spleen of hepatocytes anti-regeneration and liver injure rat,and observed the differentiation of MSC in rat.We built a experiment fundmental for treatment of livre failure by HTx.Materials and Methods:1.Human embryonic MSC were isolated by density gradient centrifugation from abortus,and cultured by MEM culture medium in vitro.MSC were detected the content and activity after isolated and cultured.2. Trial rats had been done the partial hepatectomy after they were feed by AAF.Rats were detected the hepatic function by schedule.3. MSC were transplantated into the spleen of anti-regeneration of hepatocytes and liver injure rat. We took spleens and livers from rats after operation in schedule,and had learned that the human ALB,CK 8,CK 18 and CK19 antibody expression of immuno- histochemistry staining.By staining for glycogen,we knew the content of glycogen in cells.Results: 1.There is from 85% to 93% of MSC in the isolated cells.The content of MSC is more than 93% after cultured 7 days in vitro,and the content of activity cells is more than 95%.2. After had been feed by AAF,it is obviously inhibition that the regeneration of hepatocytes in the liver injure rats.The recovery of hepatic function was slowly.3.Human CK 8,CK 18 and CK19 antigene expression were found in rats' spleens after 10 days of MSC transplantation by IHC staining.Human ALB antigene expression was also found in spleens after two weeks later of MSC transplantation.The expressions of spleens were shown in the later 12 weeks after the transplantation. The positive expressions of livers is similar.Most positive cells were in the portal area.It shows that positive cells came from spleen by portal vein.There is no positive cells in liver of control group. By staining for glycogen,we found positive cells in spleen after two weeks later,and the positive cells can be found 12 weeks after transplantation.It shows that the differentiated cells have the abilty of Synthesis glycogen.Conclusions:1. The purity of Human embryonic MSC which isolated by density gradient centrifugation and cultured by MEM culture medium in vitro is high.The activity of MSC is good.2.After had been feed by AAF,it is obviously inhibition that the regeneration of hepatocytes in the liver injure rats.It is a good animal model of hepatocytes anti- regeneration and liver injure.3. Human embryonic MSC can differentiate into hepatocyte in a inhibition of hepatocytes regenerate and liver injure rat's spleen.Part of MSC move into liver and differentiate into h...
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