The In Vitro Study Of Isolation, Culture And Identification Of Umbilical Cord Blood Mesenchymal Stem Cells

[Objective] To investigate the optimal method for the isolation , culture and identification of human umbilical cord blood mesenchymal stem cells(MSCs)in vitro ,and to observe the feasibility of using MSCs as seed cells for cell therapy.[Methods] Umbilical cord bloods(n=36) were collected from full term childbirth scheduled for cesarean and vagina deliveries. All specimens were obtained sterikly with preservative-free CPDA-1 compound natrium citricum. Half of the bags(n=18) UCB mononuclear Cells (MNCs) were isolated directly by Ficoll-Hypaque,and the MNCs were Cultured respectively by DMEM-LG with 5%, 10% and 20% fetal calf serum(FCS) medium to produce adherent layer;the other bags MNCs were isolated by indirect method:Firstly,the RBC were segregated by cellogran;Secondly, MNCs were isolated from the collected WBC by Ficoll-Hypaque densiby gradiet centrifugation,the isolated MNCs were cultured respectively by DMEM-LG with 5%, 10% and 20% FCS to produce adherent layer.Bone marrow(BM)(n=11)MNCs were isolated and cultured as control group,cells growth characteristics were observed and surface markers were detected by flow cytometry.