不同的冻存方法对脐血CD34~+细胞的影响

目的 :研究不同冻存方法对脐血CD34+ 细胞的影响 ,探讨脐血CD34+ 细胞先分离后冻存的可能性 ,以寻找脐血保存的更好方法。方法 :对 31份脐血采用全脐血直接冻存 ,分离为单个核细胞冻存 ,和分离为CD34+ 细胞冻存 ,3种不同的冻存方法 ;用造血祖细胞培养技术、活细胞染色技术、流式细胞技术等方法研究冻存前后脐血造血祖细胞活性的变化。结果 :冻存后细胞活度下降 ,集落形成率下降 ,以全脐血冻存组为甚 ;CD34+ CD38- 比例冻存后增高。结论 :先分离后冻存法可行 ,且具有减小冻存体积 ,对造血祖细胞损伤较小的特点 ;早期造血祖细胞可能更能耐受冻存处理

Recovery of CD34 Cells After Cryopreservation of Umbilical Cord Blood Cells in Three Different Methods

To study the effect of umbilical cord blood(UCB) transplantation, and combination of UCB transplantation and gene therapy, one of major therapy for hereditary hematological disease in the future.Cryopreservation of UCB was critical for the use of UCB.This experiment was to optimize the procedure of cryopreservation.Methods:Three different cryopreservation procedures-whole UCB,mononucleate cells' suspension and CD34+ cells' suspension were used in this study.Recovery of CD34+ cells was evaluated by viability assay,clonogenic capacity assay and percentage of CD34+ cell assay.Results:The viability and clonogenic capacity were decreased after cryopreservation,especially cryopreservation of whole UCB directly.The percentage of CD34+CD38- cells was increased after cryopreservation.Conclusion:①Cryopreservation after separation is feasible.If CD34+ cells are going to be separated by magnetic system,cryopreservation after separation will not only reduce the storage volume but also lead to less damage of CD34+.②The early hematopoietic progenitors might be more tolerance to the cryopreservation.