低浓度二甲基亚砜冻存外周血造血干细胞的效果

Analysis of hematopoietic stem cells cryopreserving with low concentration dimethysulfoxide

目的:观察外周血造血干细胞(PBSC)用低浓度二甲基亚砜(DMSO)和羟乙基淀粉(HES)在-80℃条件下冷冻保存的效果。方法:49例次患者的PBSC在冻存后3、6个月及1年进行复苏,分别取样进行有核细胞(NC)计数、锥虫蓝拒染率测定、CD34+细胞阳性率分析和粒-巨噬细胞集落生成单位(CFU-GM)检测。结果:PB-SC在-80℃冰箱中冻存3个月和6个月的NC、CD34+细胞、CFU-GM回收率的差异均无统计学意义(均P>0.05),NC活细胞比率差异也无统计学意义(P>0.05)。PBSC冻存1年NC、CD34+细胞、CFU-GM集落回收率及活细胞比率均有显著性下降(均P<0.01)。结论:应用低浓度DMSO在-80℃冻存PBSC3个月和6个月的细胞能得到较好保存,1年后的冻存效果显著下降。

Objective：To evaluate the cryoprotective effects of hematopoietic stem cells with low concentration dimethysulfoxide and HES. Methods The viability and function of hematopoietic stem cells from 49 patients were investigated before and after cryopreserving with low density dimethysulfoxide for three months, six months and one year. CD34 positiue rate were analyzed by flow cytometry, CFU-GM was detected in vitro, Nucleated cells （NC） were detected by microscope, the survial rate was determined by trypan blue staining. Result： The results showed the difference of recovery rate of CD34^＋ , CFU-GM , NC were significant at three different cryopreserved interval. Conclusion：PBSC can be well cryopreserved at --80℃ with low concentration dimethysulfoxide for three months and six months, but not for one year.