Isolation and biological characteristics of rat umbilical cord mesenchymal stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:There are many studies concerning rat bone marrow mesenchymal stem cells for immune tolerance following transplantation and tissue repair.However,there are no reports on umbilical cord mesenchymal stem cells(UCMSCs).OBJECTIVE:To establish a method of separating mesenchymal stem cells(MSCs)from rat umbilical cord,and to study its biological characteristics.METHODS:MSCs were separated from rat umbilical cord with enzyme method and tissue mass method,and then incubated in DMEM-LG medium.Cell morphology was observed under an inverted microscope.Growth curves of cells were drawn using cell counting.Cell cycle and surface antigen were detected with flow cytometry.Adipogenic differentiation and osteogenic differentiation were tested by immunohistochemistry.RESULTS AND CONCLUSION:Both of the two methods could obtain plenty of MSCs from rat umbilical cord.Primary culture showed that the efficiency of enzyme method was higher than tissue mass method.Passage time of the former was about 10 days and the latter was 14 days.The passage time of latter except primary culture was the same.Immunophenotype analysis showed that MSCs from rat umbilical cord expressed adhesion molecule and stromal cell markers,CD90 and CD106,but did not express hematopoietic cell markers,CD34 and CD45.In vitro induction test verified that rat UCMSCs have the potentials of adipogenic and osteogenic differentiation.