Isolation,characterization and differentiation of human umbilical cord bloodderived mesenchymal stem cells

Objective To isolate and culture human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSC),and to observe the biological characteristics of hUCB-MSC and to investigate the osteogenic and adipogenic differentiation potential with proper condition in vitro.Methods Umbilical cord blood mononuclear cells were separated by the method of density gradient centrifugation and MSCs were obtained by adhere culture.The cell morphology and their growth characteristics in vitro were observed under an inverted phase contrast microscope,and the cell cycles were tested by flowing cytometry.Mesenchymal like stem cells were identified by surface marker with flow cytometry.The osteogenic potential was determined by alizarin red staining and the adipogenic potential was detected by oil-red O staining.Results UCB-MSCs were adherent with a fibroblast like morphology and whirlpool like growth alinement.Flow cytometry showed that The third passage cells were positive for CD105,CD73 and CD90 expression,but they failed to express hematopoictic cell surface markers,such as CD34 and CD45.When the MSCs were induced to osteogenic and adipogenic differentiation for 3 weeks,the formation of a mineral extracellular matrix and neutral lipid vacuoles detected by alizarin red staining and oil-red O staining respectively.Conclusions The MSCs from umbilical cord blood in this study show great capability of proliferation and regeneration and can be induced into osteoblasts and adipoblasts in a suitable condition in vitro.