-80℃低温冻存保护剂对不同来源的造血干细胞体外冻存效果分析  

Efficacy of external cryopreservation with-80℃ cryoprotectant to hematopoietic stem cell from different sources

作　　者：吴书一[1] 熊灵军[2] 刘文刚[1] 罗明霞[3] 赛亚[3] 朱趁红[3] 董秀娟[3] 赵晓武[3] 

机构地区：[1]郑州市第三人民医院干细胞与器官移植实验室,郑州450000 [2]郑州大学第二附属医院消化疾病研究所 [3]郑州市第三人民医院血液科

出　　处：《医药论坛杂志》2011年第10期3-6,共4页Journal of Medical Forum

基　　金：河南省自然科学基金项目(0411045200)

摘　　要：目的分析-80℃体外冻存保护体系对不同来源的造血干细胞的冻存效果。方法以国产或进口药用二甲基亚砜(DMSO)、羟乙基淀粉(HES)、人血白蛋白(HSA)配制细胞冷冻保护剂,分别对人外周血、纯化的CD34+、骨髓及脐血造血干细胞进行-80℃低温保存,冻融前、后计数MNC、CD34+细胞、CFU-GM数量、回收率及台盼蓝拒染率,了解其生理活性。结果 4种来源的造血干细胞各30例,以120g/L HES加10%(V/V)DMSO和64g/L HSA为保护剂-80℃冻存360d,观察组与对照组在冻融前、后MNC、CD34+细胞、CFU-GM数量、回收率及台盼蓝拒染率无统计学意义(P>0.05)。应用国产试剂对118例外周血干细胞、39例纯化CD34+造血干细胞、3例脐血干细胞进行-80℃低温冻存1～9个月,进行冻存的造血干细胞移植,移植后7～19d内均成功恢复造血重建。结论以120g/L HES加10%(V/V)DMSO和64g/L HSA为细胞冷冻保护剂,能够良好保持干细胞数量及生理活性,且国产试剂与进口试剂无显著差别,具有价格低廉,容易购买,低毒安全,操作简便等特点,可临床应用于造血干细胞-80℃低温短期保存。Objective To analyze the effect of external cryopreservation for hematopoietic stem cells（HSCs） from different Sources with-80℃ cryoprotectant.Methods The cryoprotectant was compound of domestic or imports medicinal dimethyl sulfoxide（DMSO） ,hydroxyethyl starch（HES） and human serum albumins（HSA） .The sources of hematopoietic stem cells were from peripheral blood,bone marraw,cord blood and purified CD34＋ cell.Physiological activity of HSCs were analyzed and evaluated before and affter-80℃ freezing according to the numbers of MNC,CD34＋ cells,CFU-GM,and recovery rate,blue placenta resistent dyeing rate.Results Totally 30 cases were in this study.The cryoprotectant compost of HES（120g/L） ,plus DMSO（10%,V/V） and HSA（64g/L） .It was no significant difference（P0.05） between the observation group and the control group.The results of cell blue placenta resistent dyeing rate and the recovery rate of MNC,CD34＋ cells,CFU-GM before and after freezing-stored 360 days were analogous.The hematopoietic recovery time in 118 cases PBSCT,39 cases purified CD34＋ cell,and 3 cases cord blood were 7～19 days,using the domestic reagents.Conclusions The cryoprotectant composting of HES（120g/L） ,plus DMSO（10%,V/V） and HSA（64g/L） can keep cell livability and the recovery rate of MNC,CD34＋ cells,CFU-GM.It had no significant difference between the domestic reagents and imported reagents.It can be applied to the protection of HSCs in-80℃ preservation for short-term.

关 键 词：低温保存 造血干细胞 羟乙基淀粉 二甲基亚砜 人血白蛋白 

分 类 号：R457.7[医药卫生—治疗学]