-80℃条件下不同时间外周血干细胞冻存效果的观察

The Observation on the Effect of Preservation Time on the Activity of Peripheral Blood Stem Cells under -80℃

目的 观察外周血干细胞在-80℃条件下保存时间与细胞活性的关系.方法 采用一种简便的-80℃干细胞冻存方法,其干细胞保护剂终浓度为5%二甲基亚砜（DMSO）,3%羟乙基淀粉（HES）和4%人血白蛋白（HAS）,不经程序降温,直接-80℃冰箱冻存,并调整细胞浓度为（2～4）×107/mL,分别对冻存1、3、6、9、12个月干细胞活性进行检测,观察台盼蓝拒染率和MNC、CD34＋细胞的回收率.结果 干细胞活性在12个月内均无明显下降,其台盼蓝拒染率和MNC、CD34＋ 细胞的回收率都在80%以上.不同时间冻存的干细胞活性比较无明显统计学差异（P＞0.05）.结论 5% DMSO、3% HES 和4% HAS作为干细胞冷冻保护剂直接在-80℃冰箱冻存外周血干细胞的方法能有效地保护外周血干细胞活性.

Objective To observe the effect of preservation time on the activity of peripheral blood stem cells under -80℃.Methods PBSC were harvested and cryopreserved to -80℃ immediately with protective agent which final density was 5% DMSO,3% HES and 4% HAS.PBSC were cryopreserved at cell concentration of （2 - 4）× 107/mL,without controlled-rate freezing to -80℃.Then the cell viability was measured at the different time（1,3,6,9, 12months after cryopreservation）by excluded trypan blue,and the recovery of MNC,CD34＋ cells.Result Cell viability had no significant decline within 12 months in each group.The recovery of MNC,CD34＋ cells at the different time was above 80%,which was no significantly decreased（P 〉 0.05）.Conclusion The cryopreservation method of PBSC in -80% freezer with the protective agent is a safe and effective cryopreservation technique to keep the activity of PBSC.