Comparison of Isolation and Culture Methods between Umbilical Cord Mesenchymal Stem Cells and Umbilical Cord Blood Mesenchymal Stem Cells

Objective To establish methods for isolation and culture of umbilical cord mesenchymal stem cells(UC MSCs) and umbilical cord blood mesenchymal stem cells(UCB MSCs) in vitro,and compare the effects.Methods Umbilical cord blood(n=30) and Umbilical cord(n=30) were collected from healthy full-term deliveries.Human UC MSCs were isolated and cultured using adherence technique and enzyme digestion,and mononuclear cells were isolated from umbilical cord blood with the lymphocyte isolation method and hydroxyethyl starch sedimentation + lymphocyte isolation method to culture UCB MSCs.Cells were cultured in DMEM/F12 containing 10% fetal bovine serum,and compared cell morphologic change and isolation outcomes.The surface markers of MSCs were detected by flow cytometry,and the ability of MSCs differentiation was analyzed by inducing differentiated in vitro.Results About 8 days after adherence,the fibroblast-like cells were observed sprawled out from the edge of the tissue,growing in cluster;and 5 days after enzyme digestion,cells growing fiber shaped.In UCB MSCs isolation and culture,compared with the lymphocyte isolation method,the number of mononuclear cells significantly increased using the hydroxyethyl starch sedimentation + lymphocyte isolation method.Two sources of MSCs surface markers analyzed by flow cytometry showed that they expressed the characteristics of MSCs.Inducing differentiated MSCs in vitro showed they could be induced to osteoblast and adipocyte.Conclusion The period of UC MSCs primary culture was shorter,and with a better culture efficiency.The hydroxyethyl starch sedimentation + lymphocyte isolation method isolated and cultured UCB MSCs was more effective.