Study of peripheral blood stem cell cryopreserved at -80℃ non rate-controlled freezing
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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,120 g/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15~720 d,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 240 d 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12~46 d(平均 29 d)的干细胞,患者于干细胞回输后14~27 d(平均 20.5 d)获得造血功能重建。结论:在采用终浓度为 60 g/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(240 d内)保存,效果理想。Abstract: 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 (P>0.05). Although recovery rate of CD34+ and CFU-GM decreased obviously after cryopreservation for 240 days (P<0.05), 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.
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