Optimization of Culture Conditions for Human Umbilical Cord Blood Mesenchymal Stem Cells(UCB-MSCs)

Objective To optimize the culture conditions for the cultivation of human umbilical cord blood mesenchymal stem cells(UCB-MSCs).Methods Samples were collected from the Obstetrics and Gynecology Department of Zhujiang Hospital.The parameters examined in this study included the sample volume,the time of first medium change,age of the fetus and the type of culture medium.Expression of surface markers was analyzed by flow cytometer.Results Under the same condition,the optimum volume of cord blood sample for successful cultivation of UCB-MSCs was 10 ml.The first medium change should be performed at 96 hours after incubation,as it could facilitate the attachment of mononuclear cells.A high successful rate of UCB-MSCs cultivation was also obtained from premature infant cord blood.The quality of fetal calf serum is also important in maintaining the growth of UCB-MSCs.Flow cytometric analysis showed that UCB-MSCs strongly expressed CD13,CD29 and CD44,but not CD34,CD45,CD14 and endothelial cell marker CD106.Conclusion Mesenchymal stem cells can be obtained and cultivated from cord blood.Sample size,the time of first medium change,fetal age and culture medium are the key factors affecting the cultivation of UCB-MSCs.