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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 non-programmed cooling at -80 ℃ for preserving autologous peripheral blood stem cells (PBSC). Methods: The peripheral blood stem cells were mobilized by conventional methods. The stem cells, 120g / L hydroxyethyl starch, 10% DMSO, 10% AB plasma were used as cryoprotectants and the stem cells were separated by 1: 1 ratio of mixing, and then placed directly at -80 ℃ refrigerator. The recovery rate of mononuclear cells (MNC), CD34 + cells, granulocyte-mononuclear cell colony forming unit (CFU-GM) and the trypan blue exclusion rate of the cells before and after storage were determined. Results: The non-programmed direct cryopreservation of 15-720 d, PBSC trypan blue exclusion rate, MNC recovery was less (P> 0.05), while the CD34 + cells, CFU-GM recovery decreased after 240 d (P <0.05), but still reached 82.5% and 79.6% respectively. Twenty-one patients were transfused with stem cells stored at -80 ℃ for 12-46 days (mean 29 days). The patients received hematopoietic reconstitution from 14 to 27 days after transplantation (average 20.5 days). Conclusion: In the case of cryoprotectants composed of 60% hydroxyethyl starch, 5% DMSO and 5% AB plasma, the non-programmed cooling method at -80 ℃ is suitable for PBSC The short-term (240d) save, the effect is ideal.