Culture And Identification Of Rat Umbilical Cord Mesenchymal Stem Cells

ObjectiveMesenchymal stem cells isolated from Wharton's jelly of rat umbilical cord,which were cultured and identified in vitro,are proposing to be an alternative source of allotransplantation cell therapy research.Materials and methodRat umbilical cord mesenchymal stem (rUCMS) cells were harvested from E19 pregnant rats operated by cesarean section , which were isolated from Wharton's jelly of rat umbilical cord and vessels were removed by mechanical dispelled. rUCMS cells were maintained in DMEM/F12 medium supplemented with 10% fetal bovine serum(FBS), 100 units/ml pecicillin, and 100ug/ml streptomycin, followed with passaged, freezed and thawed. 3rd passage of rUCMS cells were conducted further multiple experimental research, including cell proliferation assay, incorporated bromodeoxyuridine(Brdu) immunocytochemistry, adippgenic and osteogenic differentiation, flow cytometry.ResultsCells derived from rat umbilical cord exhibited spindle-shape fibroblast-like morphologies and adhered-plastic characteristic that correspond with mesenchymal stem cells(MSCs). Wharton's jelly cells of rat umbilical cord contained MSC-like proliferative capacity which 92.14% staged G0-G1, mostly incorporated Brdu within 48-hour period, differentiated into osteoblasts and adipocytes, expressed molecular markers CD29,CD 44,CD90 rather than CD34,CD45. ConclusionsCells harvested from Wharton's jelly of rat umbilical cord commited to the defining criteria of MSC on morphology,proliferation, differentiation capaticity and immunophenotyping, termed rUCMSCs. rUCMSCs are multipotent stem cells, could be regarded as a new cell source for cell-based therapy and engineering research.