Study on Isolation and Culture of Mesenchymal Stem Cells from Human Umbilical Cord Blood in Vitro and Their Biological Characteristics

Objective:To establish an optimized method to isolate,culture and expend mesenchymal stem cells(MSCs)from hu- man umbilical cord blood(hUCB)in vitro.Methods:The mononuclear cell was isolated by density gradient centrifugation with lympho- cyte separation medium from hUCB collected from full term deliveries scheduled for cesarean section.The MSCs were purified by adhe- sion with acidized D-MEM/F-12 medium.After that,the growth curves of MSCs in different passages were drawn by cell counting,and the expressions of CD29,CD105,CD34,HLA-DR in MSCs was detected by flow cytometry.Results:The adhesion cells could be ob- tained successfully and reliably from hUCB.The cells displayed a fibroblast-like morphology,and several MSCs-related antigens such as CD29 and CD105 were expressed,while neither CD34 nor HLA-DR could be found on the cells by FACS.Conclusion:MSCs could be obtained from hUCB with the method mentioned in this article and hUCB could be an alternative source of MSCs for experimental and clinical applications.