| IntroductionAs is known to us, the use of liver transplantation is limited because of the shortage of the donor. With the development in isolation of hepatocyte, it became possible to put hepatocyte transplantation(HCT) in practice. Some animal and clinical research has shown that hepatocyte can maintain its structure and express sorts of biological functions such as synthesizing the protein, metabolism of bilirubin, reservoir of hepatin after transplanted in liver, spleen or some other organs. So it is a promising way to cure acute hepatic failure, terminal hepatic disease and metabolic hepatic disease using HCT instead of liver transplantation.There are some difficulty in using of hepatocyte transplantation such as the shortage of normal human hepatocyte and as for fetal hepatocyte it is ethic and source problems. For these reasons, hepatocytes isolated from the liver of a normal male donor were transfected with Simian Virus 40 large T(SV40LTag) by Dr Li of our research group.One of the resulting hepatocyte has shown the immortalized characteristics in vitro. In some examination, it manifests highly differentiated liver function and retains the morphological characteristics of normal human hepatocyte.In this research, we have transplanted this new hepatocyte line from human into the spleen of BALB/c nake mouse to clarify its morphological and biological characteristics in vivo.Materials and MethodsBALB/c naked mice were randomly divided into two groups. Group A were removed of left liver lobe and then transplanted human-derived hepatocytes in the spleen. Group B were transplanted human-derived hepatocytes in spleen without the removal of left liver lobe. 1, 3, 7, 15and 30 days after transplantation, the regeneration of the liver was examined. Tissue sections were made from the spleens, then pathological research and immunohistochemistry were performed to study the distribution and multiplication of human-derived hepatocytes in the spleen. Expression of glycogen, AFP, ALB and HEP were detected. Then tissue sections made from heart, liver, lung, kidney and omentumwere stained by HE and immunohistochemical method to detect the migration of human-derived hepatocytes.ResultsFor Group A, 7 days after the transplantation, the volume of the liver looked like that before the resection. 15 days after the transplantation, the liver became much larger than that before the resection. The splenic tissue sections stained by HE displayed that from the first day to the 30th day, human-derived hepatocytes grew among the splenic nodules for both two groups. These hepatocytes were rich in cytoplasma and larger than the spleen cells around. According to the results of immunohistochemistry, the hepatocytes expressed HEP, a kind of specific marker of hepatocyte. However, expression of AFP and ALB were not detected. The immunohistochemical staining of K.i-67 indicated human-derived hepatocytes proliferated actively after the transplantation and reached the climax on the 3rd day for Group A, while the proliferation became weak 30 days after the transplantation for Group B. Staining of PAS showed that within 3 days after transplantation, the glycogen reserved in human-derived hepatocytes was at a low level,but it increased 7 days later. The immunohistochemical staining of Ki-67 showed that human-derived hepatocytes moved into hepatic parenchymal and maintained active multiplication from the 1st day to the 30th day. HE staining and immunohistochemical staining of HEP showed human-derived hepatocytes did not migrate to heart, lung, kidney and mesentery for both groups.ConclusionsThat is the first animal experiment about the transplantation of human-derived hepatocytes into BALB/c naked mice in our country. The human-derived hepatocytes could survive long after the transplantation. They proliferated and expressed HEP, a kind of specific marker of hepatocytes. About 7 days after the transplantation, the hepatocytes restore glycogen reserve, without the expression of AFP and ALB. The hepatocytes, which...
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