Rat Umbilical Cord Mesenchymal Stem Cells On Liver Cell Proliferation, Apoptosis And Function

ObjectiveTo investigate the effect of liver failure serum on the CD86of UCMSC. Tounderstand UMSCs on rat primary hepatocytes proliferation, apoptosis and secretoryfunctions.MethodsMesenchymal stem cells were isolated from rat umbilical cord by collagenase andtrypsin. Observe cell morphology and growth status.Cell surface markers of CD90,CD105of UCMSC were analyzed by flow cytometry method. The3rd generation of UMSCs at70%~80%fusion were randomly assigned to3groups: control group(normal culturemedium), normal serum medium group(including10%normal rat serum culture medium),serum liver failure group (containing10%serum medium liver failure). The expression ofCD86was assessed by flow cytometry after24hs. Discrete liver cells were isolated by twosteps collagenase perfusion via portal vein. Liver cells co-cultured with UMSCs withTranswell. The primary rat hepatocyte were divides into three groups: The UMSCs group,the primary rat hepatocyte group and UMSCs co-culture with the primary rat hepatocyte.The cells proliferation rate of every group were measured by MTT on1d,3d,5d,7d. LPSinduced liver cell apoptosis, and the apoptosis rate of every group were measured by flowcytometry after24hs. The concentration of albumin in the culture medium were measuredby ELISAon1d,3d,5d.Results1. UMSCs were harvested successfully from UC by collagenase and trypsin. Adherentcells were observed after24hours. As the passage number increased, cells appeared asfusiform, UMSC proliferated quickly. Flow cytometric analysis showed that the UMSCsexpressed CD90,CD105, and the rate was99.7%,99.5%.The expression of CD86on the surface of UMSCs in each groups was cultivated24h was0.5%，0.6%，0.4%.（P>0.05）2.(1) OD value of the UMSCs co-culture with primaryrat hepatocyte group was significantly higher than the OD values of UMSCs group and theprimary rat hepatocyte group on1d,3d,5d,7d（P<0.05）(2) After induction of apoptosisby flow cytometry liver cells, the positive control group, negative control and UMSCs with1*104/ml,1*106/ml co-culture with primary rat hepatocyte group of apoptosis rate was93.3%,54.2%,59.7%,72.9%,(P <0.05).(3) UMSCs didn’t secrete albumin, theconcentration of albumin in the culture medium of the UMSCs co-culture with primary rathepatocyte group was higher than the primary rat hepatocyte (P <0.05).ConclusionsThe trypsin and collagenase digestion procedure is a easy, convenient and reliablemethod to obtain UMSCs.The UMSCs expressed CD90,CD105.Serum liver failure afterintervention training UMSCs CD86expression remains.UMSCs can promoteproliferation,inhibition of apoptosisof primary rat hepatocyte and increase secretion ofalbumin.