| Mesenchymal stem cells(MSCs) are multipotential stem cells derived from mesoderm of early phase embryo. It possesses wide application perspective for inducing immunotolerance in organ transplantation. Now MSCs are mainly derived from the adult bone marrow., but many studies showed that human umbilical cord was the better sorce of mesenchymal stem cells. There are many studies that MSCs derived from rat bone marrow were used for transplantation immune tolerance and tissue repair. But because of no correlated study for MSCs derived from rat umbilical cord, study for MSCs derived from umbilical cord could not be done in vivo. In our study, we isolated MSCs from rat umbilical cord and studied their biological characteristics. At last we find a suitable method for isolating and culturing Mscs. We did experiments in vitro and in vivo to study thei effect for inducing immunotolerance and softening immunological rejection.In first part of our study, we successfully isolated MSCs from rat umbilical cord by two enzyme method and tissue mass method.Then we analyzed the cells phenotype and induced cell differentiation Results showed that adherent cells obtained through two methods were all MSCs. Their biological character were the same with MSCs'form bone marrow reported in literature. Two enzyme method was better then tissue mass method. In second part of our study, we choosed Lewis rat's spleen cells as react cells and DA rat's spleen cells as stimulate cells. Then MSCs form Wistar rat's umbilical cord were added in them. Methods By the means of 3H-TdR (three tritium thymine deoxyribosid), we do the MLC(mixed lymphocyte culture) experiments to detect whether the MSCs from rat umbilical cord affected MLC. Results showed that MSCs from rat umbilical cord could not cause rats lymphproliferation response, but have a notable suppressing effect in MLC. In third part of our study, we choosed DA rat as donor and lewis rat as recipient, to establish a heterotopic model of rat heart transpalntation and after orperation immediately inject MSCs from Wistar rat umbilical cord. We investigated the influence of MSCs on allograft of rat heart transplantation In this part,all recepients were divided into two groups. In control group,recepients received normal saline via tail vein after transplantation. In experimental group Recepients received third party MSCs via tail vein. Results showed that the surviving time of transplanted hearts in control group and in experimental group was 7.5±0.5 days and 14.8±2.1 days. Statistical analysis showed that it was longer in experimental group than that in control group(p<0.05).Our research showed that we could get a plenty of MSCs from rat umbilical cord through two enzyme method and tissue mass method. Two enzyme method was better then tissue mass method. As a third party cell, MScs from umbilical cord could suppress lymphocyte proliferation in vitro, and prolong xenograft survival and soften immunological rejection in vivo. |
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