Improved method for isolation and culture of mesenchymal stem cells from human umbilical cord blood

Objective To improve the method for isolation and culture of human umbilical cord blood-derived mesenchymal stem cells(hUCB-MSCs) in vitro.Methods The cord blood mononuclear cells were isolated by density gradient centrifugation and directly adherence growth method.After cultured with DMEM containing low glucose and 15% FBS medium for 3 passages,surface antigen expression of MSCs was detected by flow cytometry.Results The cord blood mononuclear cells began to adherent to flasks in 24h,mainly displayed osteoblast-like cells and mesenchymal-like cells.By natural purification for 3 passages,long spindle shaped-MSCs were harvested.Test results from flow cytometry showed that the cells expressed CD44 strongly.Conclusion With the improved method hUCB-MSCs can be isolated,cultured and expanded in vitro.The harvested hUCB-MSCs express the identical surface antigen as human bone marrow-derived mesenchymal stem cells(BM-MSCs).