人自体血清支持脐血源HSCs和MSCs共培养研究

Co-culture of HSCs and MSCs derived from umbilical cord blood using human autologous serum

考察使用cytodex-3微载体共培养人脐带血(UCB)来源的造血干细胞(HSCs)和间充质干细胞(MSCs)的可行性,并评价其应用于未来临床治疗中的可能性.考虑到临床应用的安全性要求,尝试采用采集于UCB的人自体血清(HAS)取代目前广泛使用的胎牛血清(FBS),以期在体外共培养后可同时获得UCB-HSCs和UCB-MSCs.由于MSCs贴附的cytodex-3载体与悬浮的HSCs之间的密度差异,在共培养后自由沉降分离这两种不同类型的干细胞将会十分容易.为优化培养条件,评估了用不同含量的HAS(2.8%、5.6%、8.3%和11.1%)对UCB-MSCs和UCB-HSCs扩增的影响.结果表明,5.6%HAS培养UCB-HSCs最优,扩增倍数达(1.88±0.33)倍,且同时对UCB-MSCs的扩增亦无很大影响.因此,共培养条件下的HAS最佳含量约为5.6%.所研发的共培养体系可为临床应用提供扩增的UCB-HSCs和UCB-MSCs.

The feasibil cells （MSCs） derived ity of co-culturing hematopoietic stem cells （HSCs） and mesenchymal stem from human umbilical cord blood （UCB） using cytodex-3 microcarriers is investigated in order to assess it as a possibility for future clinical therapies. Considering the safety requirements of clinical applications, human autologous serum （HAS） collected from UCB is used to replace the widely-used fetal bovine serum （FBS）. Following to this, after UCB-HSCs and UCB- MSCs ex vivo culture, both can be harvested simultaneously. Besides, after their co-cuhure, the two different types of stem cells can be easily separated by sedimentation due to the density differences between cytodex-3 microcarriers （containing adherent MSCs） and the suspended HSCs. In order to optimize the culture conditions, the effect of the content of HAS （2.8%, 5.6%, 8.3% and 11.1%） on the expansion of UCB-MSCs and UCB-HSCs is assessed. The results indicate that the expansion of UCB-HSCs is at least （1.88±0.33）fold while supplied with 5.6% HAS, better than that in the other groups, while it has little effect on the expansion of UCB-MSCs. It is concluded that the optimal content of human autologous serum for the co-culture conditions herein reported is about 5. 6%. Finally, the co-culture system developed and herein reported is feasible to provide expanded UCB- HSCs and UCB-MSCs for clinical applications.