人脐血间充质干细胞的培养条件比较

目的摸索人脐血间充质干细胞（mesenchymal stem cell,MSC）的培养条件。方法根据不同采血量、首次换液时间、胎龄、不同培养基对样本分组，比较不同培养条件对脐血中的间充质干细胞原代生长的影响，以流式细胞仪对培养出的间充质干细胞进行细胞表面标志检测。结果在相同条件下，取10ml的脐血能较大程度培养出间充质干细胞；观察首次换液时间在培养后96h较为合适，延长换液时间有利于数量不占优势的单核细胞充分贴壁：早产胎儿的脐血培养出的间充质干细胞成功率较高；胎牛血清的质和量决定了培养成功与否。培养出的间充质干细胞不表达造血细胞系的标志（CD34、CD45、CD14）及内皮细胞的标志（CD106），强表达CD29、CD44、CD13。结论样本量、首次换液时间、胎龄、培养基的质和量对MSCs的成活、生长有关键作用。

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 l0 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.