实用细胞冻存技术在自体造血干细胞移植中的应用

Application of practical cell cryopreservation technique in autologous hematopoietic stem cell transplantation

目的:分析实用细胞冻存技术在自体造血干细胞移植中的应用。方法:收集11例患者自体造血干细胞移植(AHSCT)动员的外周血造血干细胞,收集袋总容量100~150 m L,其中二甲基砜(DMSO)占总容量的10%、异体同型病毒灭活血浆占20%、外周血造血干细胞占70%。超净台无菌操作分装后放置于-20℃的冰箱冻存12 h,再置于-80℃冰箱冻存。与常用冻存技术做对照,移植前复苏干细胞,检测细胞活性、单个核细胞计数(MNC)和CD34+细胞的回收率;移植后观察患者粒细胞、血小板植入时间。结果:复苏干细胞后检测细胞活性(89.10±3.98)%,MNC和CD34+细胞的回收率分别为(91.84%±2.32)%和(90.59%±1.72)%,与对照组相比,差异无统计学意义(P>0.05);移植后粒细胞中位植入时间+12 d、血小板中位植入时间+18 d,与对照组相比,差异无统计学意义(P>0.05)。结论:DMSO与病毒灭活血浆是一种经济、易操作的干细胞冷冻保护剂。

Objective:To analyze the application of practical cell cryopreservation technique in autologous hematopoietic stem celltransplantation.Methods:The mobilized peripheral blood stem cells of11patients with autologous hematopoietic stem celltransplantation(AHSCT)were selected.The total volume of collecting bag was100～150mL;the two methyl sulfone(DMSO)accounted for10%of total volume;the allogeneic virus inactivated plasma accounted for20%;the peripheral blood stem cellaccounted for70%.They were placed-20℃refrigerator for freezer12hours after ultra clean aseptic operation,placed-80℃refrigerator for freezer.Compared with the commonly used cryopreservation technique,the stem cells were given recovery beforetransplantation,and the cell activity,mononuclear cell count(MNC)and CD34+cell percent recovery were detected.Aftertransplantation,the granulocyte and platelet implantation time of patients were observed.Results:After recovery stem cells,the cellactivity was(89.10±3.98)%;MNC and CD34+cell percent recovery were(91.84%±2.32)%and(90.59%±1.72)%;there was nosignificant difference that compared with the control group(P>0.05).After transplantation,the granulocyte median implantationtime was+12d;the platelet median implantation time was+18d;there was no significant difference that compared with the controlgroup(P>0.05).Conclusion:DMSO and virus inactivated plasma is an economical and easy to use stem cell cryoprotectant.