Phenotypic and Proliferative Features of Human Umbilical Cord Mesenchymal Stem Cells in Vitro

Objective To establish an in vitro method to culture stem cells derived from human umbilical cord and study the biological characteristics and differentiation potentials of this mesenchymal stem cells.Methods The fresh human umbilical cord was made into cell suspension and cultured in DMEM/F-12 mixed medium which contained l0%fetal bovine serum.The adhesion and proliferation of primary cells were dynamically observed.The P3 hUC-MSCs populations were collected,then the cellular proliferation capacity was detected by thiazolyl blue tetrazolium bromide(MTT) assay.The molecular markers(CD29,CD44,CD31,CD45 and CD34)on the cell surface were identified by flow cytometry(FCM).Oil red O stains assay was to confirm the multilineage potentials of hUC-MSCs.Results The hUC-MSCs isolated from human umbilical cord stromas exhibited fibroblastie morphology and they were positive for CD29(98.3%),CD44(99.2%),and negative for hematopoietic stem cells surface markers CD34(0.3%),CD31(0.8%) and CD45(0.4%).To induce the differentiation of hUC-MSCs toward adiposity was to confirm differentiation potentials of this cultured mesenchymal stem cells.Conclusion The hUC-MSCs are stably isolated and cultured in vitro and confirmed to be differentiated.