人脐带血全血冷冻干燥保存的初步研究  

Approach to freeze-drying wholly human navel cord blood

作　　者：习德成[1] 陶乐仁[1] 肖鑫[1] 李军[1] 华泽钊[1] 陈迪[1] 张喻[1] 

机构地区：[1]上海理工大学低温生物医学技术研究所,上海200093

出　　处：《真空》2006年第2期44-47,共4页Vacuum

基　　金：上海市教委发展基金曙光计划项目

摘　　要：脐带血是一重要的造血干/祖细胞(HSPC)来源,有着极为广泛的应用前景。目前脐带血低温保存的方法因为成本高昂而得不到有效推广,但将冷冻干燥方法运用到脐带血的保存上则可能解决这一瓶颈问题。将样品预冻到-35℃后,一次干燥时搁板温度控制在-30℃,二次干燥时搁板温度控制在+15℃,整个冻干过程在25小时的工艺下,通过改变脐带血中冻干保护剂组分及浓度,比较冷冻干燥后有核细胞的恢复率,得出恢复率最高(68.397%)的配方(40%PVP+30%海藻糖+10%甘露醇),并比较了细胞冻干前后的形态,然后将样本在常温下放置三周后进行了P I染色活性检测(89.08%)。Human navel cord blood is an important origin of HSPC. It has a broad perspect in application. At present, its applications can＇t be more widespread because the expenses of its cryogenic preservation is too high. However, the freeze-drying process provides a new way to solve this problem. The process was conducted the way a sample is precooled to -30°C, and the shelf temperature during primary drying is set to -30°C, which during secondary drying is set to 15°C. The time required for the whole freeze-drying process is about 25 hours. By changing the components and concentrations of cryoprotector agent （CPA）, the restoration ratios of nucleated ceils after drying are compared, and the highest restoration ratio is 68. 397% of which the prescription is 40% pvp＋30% trehalose＋10% mannite. The morphologies of nucleated ceils after drying are compared with the fresh ones. Then, the samples are preserved for 3 weeks for its livability detection by Flow Cytometer, of which the result is 89.08%.

关 键 词：人脐带血 冷冻干燥 冻干保护剂 活性检测 

分 类 号：R318.52[医药卫生—生物医学工程] TK124[医药卫生—基础医学]