脐血造血细胞的分离冻存  

A study on the separation and cryopreservation or cord blood hematopoietic cells

作　　者：兰炯采[1] 甘茂州[2] 陈强[3] 孟庆宝[1] 张志梅[1] 

机构地区：[1]第一军医大学南方医院输血科,广州510515 [2]四川省人民医院血液科 [3]中国医学科学院输血研究所

出　　处：《临床输血与检验》2001年第2期8-11,共4页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：四川省卫生厅;科委资助的脐血干细胞中心项目

摘　　要：目的 探讨不同条件对脐血造血细胞回收的影响。方法 运用６％羟乙基淀粉（ＨＥＳ）沉降法分离脐血，以程序降温法冻存脐血造血细胞。结果ＨＥＳ两次离心法分离脐血，造血细胞回收率高，活力好，费时短。脐血分离前在４°Ｃ或室温下可放置２４小时而不影响细胞活力。ＨＥＳ终浓度以１％～２％为宜。以０．９％生理盐水＋ＤＭＳＯ做保护剂对脐血造血细胞损伤小。复苏后脐血造血细胞可放置６０分钟，洗涤会造成约１０％的细胞损失，但能延长放置时间达１２０分钟以上。结论ＨＥＳ两次离心法分离脐血简便实用。Objective To study the effect of various circumstances on recovery rate of cord blood hematopoietic cells. Methods Human cord blood has been used as an alternative source of hematopoietic cells for transplantation. We collected 266 cord blood at the time of delivery ,obtained a mean volue of (85± 25. 9)ml,and studied the effect of storage after collection, cell separation and cryopreservation under various condition. Results Red cells were sedimented by ad-dition of 1% ~ 2% HES with gentle centrifuge. Recovery rate of nucleated cells. CD34+ cells and CFU were 85. 7%, 86.4% and 86.5% respectively (n= 56). After frozen for 1 month, the recovery rate of nucleated cells and CFU were 85.6% and 91. 1% respectively (n=7). It suggested that collecting of cord blood should at 4C or room temperature for less than 24 hours, HES should be used at the concentration of more than 1% for efficient cell separation. Conclusion The method of two centrifuge was more simple and convenient than nature sediment.

关 键 词：HES 脐血造血细胞 冻存 分离 放置 复苏后 血分 DMSO 细胞活力 保护剂 

分 类 号：R556.5[医药卫生—血液循环系统疾病] R457.12[医药卫生—内科学]