脐血CD34^＋细胞分离方法比较.pdfVIP

· 6· 交通医学2008年第22卷第 1期 Med J of Communications，2008，Vo1．22．No．1 [文章编号]1006—2440(2008)Ol—OOO6—03 脐血CD34+细胞分离方法比较 李俊宏 刘 红 林赠华 姜胜华 朱云鸿 (南通大学附属医院血液科，江苏226001) 摘 要 目的：比较不同方法分离脐血单个核细胞(MNC)及纯化CD34 细胞效果。方法：采集脐血48份，在 室温下保存 12、24、48小时，分别用密度离心法和羟乙基淀粉(HES)沉淀法分离脐血MNC，再用免疫磁珠法纯化 CD34+细胞，以台盼蓝拒染法检测细胞存活率。结果：脐血在室温下保存48小时分离MNC和CD34 细胞数最多， 其次为12小时，在24小时分离所得最低(P0．05)；HES沉淀法较密度离心法分离所得MNC和CD34 细胞数明显 增多(P0．05)；6组细胞存活率差异无统计学意义(P0．05)；免疫磁珠法纯化CD34 细胞可达(88+_5)％的纯度。结 论：HES沉淀法优于密度离心法，且48小时分离为优，免疫磁珠法纯化CD34 细胞可提高纯度：各种方法对细胞 存活率无明显影响。 关键词 胎血；CD34 ；造血干细胞；细胞分离 中图分类号 Q462 文献标识码 A Comparison on Methods of Separating CD34 Cells from Umbilical Cord Blood LI Junhong，LIU Hong，LIN Zenghua，et al(Department of Hematology，Affiliated Hospital of Nantong University，Jiangsu 226001) Abstract Objective：To compare the effects of separating mononuclear cells and purifying CD34 cells with different methods． Methods：Collecting 48 cord blood units，separating mononuclear cells from cord blood by density centrifugal method and hydroxyethyl starch precipitation method after being stored for 12h，24 h and 48h at room temperature(RT)，purifying CD34 cells with immunomag- netic beads，and assessing cell viability by typan blue staining．ResuIts：The numbers of MNC and CD34 cells were the largest when being separated after being stored for 48h at RT，smaller after being stored for 12h，and the smallest after being stored for 24h(P 0．05)；more MNC and CD34 cells could be separated by HES precipitation method than by density centrifugal method(PO．05)；there WaS no significant difference between the cell survival rates of the six groups(／90．05)；the purity of CD34 cells could get to(88+_5)％ after being purified by immunomagnetic beads．Conclusions：Different separation methods have different effects；purifying CD34 cells with immunomag