Therapeutic Effect Of Transplantation Of Human Amniotic Membrane-and Umbilical Cord-Derived Mesenchymal Stem Cells On Hepatic Cirrhosis In Rats

AIMSTo evaluate therapeutic effects of human amniotic membrane-derived mesenchymal stem cells (hAM-MSCs) and human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) transplanted on the hepatic cirrhosis of rat induced by CC14.METHODSHuman umbilical cord-derived mesenchymal stem cells and human amniotic membrane-derived mesenchymal stem cells were isolated from human and cultured, then the surface makers of stem cells were analyzed by flow cytometry. Animal model of hepatic cirrhosis were induced by CC14. In8weeks, five rats were killed for pathology comformation, and thirty rats with hepatic cirrhosis were randomly divided into3groups:hAM-MSCs group(n=10), hUC-MSCs group(n=10) and control group(n=10).2×106MSCs in2ml of saline was infused via tail vein, control group was only given same volume of saline infusion. Liver function was examined before and after stem cell transplantation at4weeks. HE staining and Masson dyeing were performed. The expression level of alpha-smooth muscle actin (α-SMA) in the liver was determined by immunohistochemistry.RESULTS1. hUC-MSCs were isolated and cultured from the human umbilical cord by tissue cultivation. A large number of stem cells crawled from the human umbilical cord on the seven day under inversed microscope. After a week, lots of long spindle or flat fibroblast-like cells with whirlpool or parallel around mircro-mass were presented. hAM-MSCs were isolated from human amnion treated with trypsin and collagenase. During the primary cell cultures, the attached cells stretched after24～48h and took the shape of rond and a typical spindle-shaped fibroblast phenotype. Both isolated human umbilical cord mesenchymal stem cells and human amnion mesenchymal stem cells expressed CD29and CD44, but without CD34expression.2. After cell transplantation4weeks, liver function parameters were markedly ameliorated (ALT:204.6±16.4U/L,195.6±21.2U/L vs539.8±36.2U/L; AST180.1±25.2U/L,167.5±19.0U/L vs337.4±23.4U/L, all P<0.05).3. Combined with HE staining, we found that the degeneration and necrosis of liver cells and the inflammatory infiltration of the portal area were alleviated after cell transplantation. A reduction of pseudolobule and the hepatic fiber bundles in the rat liver after cell transplantation were observed through Masson staining.4. The expression of α-SMA was reduced in the hUC-MSCs group and hAM-MSCs group compared with control group (130.6±3.0,127.0±2.6vs152.2±5.4, all P<0.05)5. There was no statistically significant difference with the liver function and a-SMA expression between hUC-MSCs group and hAM-MSCs group after cell transplantation.Conclusion1. We were able to harvest a lot of hUC-MSCs and hAM-MSCs by tissue cultivation and by trypsin and collagenase respectively, their morphology and immunophenotype were similar to Bone Marrow MSCs.2. In rat model of hepatic cirrhosis with hUC-MSCs and hAM-MSCs transplantation, improved liver function was detected, and the degree of liver cirrhosis was reduced.3. hUC-MSCs and hAM-MSCs had the similar therapeutic effect on the hepatic cirrhosis of rat in our study, they may benefit the treatment of hepatic cirrhosis.