Therapeutic Effects And Pathways Of Human Umbilical Cord Mesenchymal Stem Cells Transplantation On Acute Liver Failure Model

Objective: To explore treatment effect and approaches of humanumbilical cord mesenchymal stem cells transplantation on acute hepaticfailure in rats.Methods: hUCMSCs were isolated from umbilical cord withattachment culture method，and the surface antigens were tested by flowcytometry. Forty-eight male or female Sprague Dawley rats (weight:180-220g, age:6-8weeks）were randomly divided into4groups (n=12):control group, model group, caudal vein transplantation group and hepaticlobe injection transplantation group. Animal model of acute liver failurewere induced by injecting intraperitoneally with50%olive oil solution ofcarbon tetrachloride2.5mL·kg-1, then treatment groups were injected withhuman umbilical cord blood mesenchymal stem cell suspension separatelythrough tail vein or injection into the liver24h post-model. Collected bloodserum and liver tissues in several times to analyze the improvement of histological repair and liver function. Real-time PCR were used to detectthe expression of human CK8、CK18and AFP mRNA in liver tissues.Immunohistochemistry were used to detect the expression of human CK18mRNA in liver tissues.Results:1. We can isolate hUCMSCs from the tissues of umbilical cord with theadherence methods and proliferate it in vitro. Expression of MSCsassociated surface antigens was detected by flow cytometry.2. There were statistically significant differences about liver functionsuch as ALT and TBil after transplantation (P<0.05). hUCMSCs improvedhistological status through enhancing hepatocellular regeneration andreducing inflammatory cells.3. Real-time PCR results showed that the expression of CK8、CK18andAFP mRNA were obviously increased in the tail vein transplantation groupand hepatic lobe injection transplantation group compared with the modelgroup (P<0.05).4. Immunochemistry results revealed that transplanted hUCMSCs inanimal liver can differentiate into functional hepatocyte-like cells thatexpressed human CK18as hepatocyte-specific marker in the caudal veintransplantation group and hepatic lobe injection transplantation group.Cells differentiated from human umbilical cord stem cells in rat liver weregradually migrated from the portal area to the central vein region. 5. No significant differences of histological repair and grade ofdifferentiation were examined between the treatment through tail veingroup or direct injection into liver group(P>0.05).Conclusion:1. hUCMSCs can be isolated and cultured successfully with adherenceculture method.2. hUCMSCs can prompt the repair of acute liver failure and enhancepathological repair after transplanting into acute liver failure rats.3. Transplanted cells in animal liver can differentiate into functionalhepatocyte-like cells that expressing hepatocyte-specific markers.4. Compared to direct injection into liver group, transplant via tail veinsgroup had similar therapeutic effects but more convenient to applicationand promotion.