-80℃非程控降温保存自体外周血干细胞的应用  

Study of peripheral blood stem cell cryopreserved at -80℃ non rate-controlled freezing

作　　者：刘淑均[1] 杨鹏[1] 王莉[2] 张循善[1] 卞茂红[1] 

机构地区：[1]安徽医科大学第一附属医院输血科,合肥230022 [2]安徽医学高等专科学校计算机教研室

出　　处：《临床血液学杂志（输血与检验）》2012年第3期345-346,349,共3页Journal of Clinical Hematology(Blood Transfusion & Laboratory Medicine)

摘　　要：目的:观察-80℃非程控降温保存自体外周血干细胞(PBSC)的应用效果。方法:常规方法动员外周血干细胞,血细胞分离机分离采集干细胞,120g/L羟乙基淀粉、体积分数为10%的DMSO、体积分数为10%的AB型血浆为冷冻保护剂与干细胞以1∶1的比例混合,然后直接将其置于-80℃冰箱保存。测定保存前以及保存后不同时间的单个核细胞(MNC)、CD34+细胞、粒-单核细胞系集落形成单位(CFU-GM)的回收率以及细胞的台盼蓝拒染率。结果:经非程控直接冷冻保存15～720d,PBSC的台盼蓝拒染率、MNC回收率的变化较小(P>0.05),而CD34+细胞、CFU-GM回收率在冻存240d后下降较明显(P<0.05),但仍分别达82.5%、79.6%。21例患者经相应处理后,回输经-80℃下保存12～46d(平均29d)的干细胞,患者于干细胞回输后14～27d(平均20.5d)获得造血功能重建。结论:在采用终浓度为60g/L羟乙基淀粉、体积分数为5%的DMSO及体积分数为5%的AB型血浆组成的冷冻保护剂情况下,-80℃非程控降温法适于PBSC的短期(240d内)保存,效果理想。Objective：To observe the effect of the cryopreservation of human peripheral blood stem cells（PBSC） by -80℃ non rate-controlled freezing. Method：We mobilized and collected peripheral blood stem in common methods ,froze cells with 120 g/L HES and 10% DMSO and 10% human plasma of AB type without rate-controlled and preserved them at -80℃.Typeran blue viability and recovery rate of mononuclear cells （MNC）, CD34＋cells, colony-forming unit-granulocyte macrophages（CFU-GM） were detected before and after cryopreservation in different periods.Result：Having been cryopreserved from 15 to 720 days at -80℃, there were no statistically significant differences in trypan blue viability and MNC recovery rate （P0.05）. Although recovery rate of CD34＋ and CFU-GM decreased obviously after cryopreservation for 240 days （P0.05）, the recovery rate was 82.5% and 79.6%, respectively.After corresponding pretreatment, twenty one patients received transfusion with -80℃ cryopreserved PBSC which had been stored for 12 to 46 days （mean 29 days） and hematological reconstitution was obtained successfully in 14 to 27 days （mean 20.5days）.Conclusion：-80℃ non rate-controlled freezing method could be suitable to human PBSC short-term cryopreservation with the cryoprotectant composed of 60 g/L HES and 5% DMSO and 5% human plasma of AB type.

关 键 词：外周血干细胞 非程控降温 冷冻保存 

分 类 号：R457.7[医药卫生—治疗学]