-80℃冰箱长期冻存外周血造血干细胞研究

Long-term cryopreservation of peripheral blood stem cells at -80 ℃

目的对外周血造血干细胞活性在深低温(-80℃)下保存3年的效果进行评价。方法对30例经MCS+血细胞分离机采集所得的外周血造血干细胞加入细胞冻存保护剂分别予以-196℃液氮冻存和-80℃冰箱深低温冻存,并检测和比较冻存前后单个核细胞(mononuclear cells,MNC)、CD34+细胞的计数及细胞回收率,细胞活性。结果 -80℃冰箱冻存组MNC、CD34+细胞冻存3年与冻存前比较差异均无统计学意义冻存3年后MNC回收率与冻存1年后比较差异无统计学意义。3年后-80℃冰箱冻存组干细胞与-196℃液氮冻存组比较,差异无统计学意义。结论 -80℃冰箱深低温保存法快速、简便、安全、费用低廉,可有效用于外周血造血干细胞长期保存。

Objective To evaluate long-term cryopreservation of peripheral blood stem cells at- 80 ℃. Methods The peripheral blood stem cells w ere collected by MCS ＋ cell separator from 30 subjects. The cells w ere preserved in refrigerator（- 80 ℃） or in liquid nitrogen（- 196 ℃） w ith cryoprotectant agent for 3 years. The count and recovery rate of mononuclear cells（ MNC） and CD34＋cells, trypan blue resistance（ TBR） w ere compared before and after cell freezing. Results There were no significant differences in the count,the recovery rate of MNC and CD34＋cells and cell viability before and after 3-year cryopreservation at- 80 ℃,betw een 3-year and 1-year cryopreservation at- 80 ℃ and betw een 3-year cryopreservation at- 80 ℃ and- 196 ℃. Conclusion- 80 ℃ freezing temperature can be used for long-term cryopreservation of peripheral blood stem cells w ith high cell viability and low cost compared to- 196 ℃ liquid nitrogen preservation.