体外培养扩增的人脐血CD34^+细胞形成HPP-CFC的能力  

Ability of in vitro amplified CD34^+ cells from human umbilical cord blood to form HPP-CFC

作　　者：李秀森[1] 郭子宽[1] 刘晓丹 侯春梅[1] 张毅[1] 郭宁[1] 毛宁[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学科学院院刊》2000年第4期285-288,291,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金! (39370 2 72 )资助课题

摘　　要：目的 :评价CD34 +细胞体外扩增时高增殖潜能集落形成细胞 (HPP CFC)的变化。方法 :利用mini MACS分离脐血CD34 +细胞 ,流式细胞仪 (FACS)测定细胞纯度 ( 86%～ 95% )。应用含rhSCF、rhIL 3、rhIL 6和rhGM CSF的培养体系扩增CD34 +细胞。分别于培养的第 1,2和 4周收取细胞进行HHP CFC测定 ,测试体系为含重组人SCF、GM CSF、IL 3、IL 6、EPO等细胞因子的半固体甲基纤维素。结果 :测试体系中加入单个细胞因子不能产生HPP CFC ,含rhIL 3和rhGM CSF体系产生的集落小且主要为巨噬细胞集落 ,由所有因子组成的体系产生的集落大而致密 ,包含粒系 3种类型。此外 ,尽管细胞总数显著增加 ,扩增的CD34 +细胞HPP CFC形成率随培养时间的延长而减少。结论 :本方法显示 ,CD34 +细胞体外扩增中可以保持HPP CFC数量。Objective:To evaluate the changes of colony forming cells with high proliferative potential (HPP CFC) in human CD34 + cell culture after in vitro amplification.Methods:CD34 + cells were obtained from umbilical cord blood by mini MACS immunomagnetic selection and the cell purity was 86 95% measured by flow cytometry analysis. The isolated cells were incubated for 4 weeks in the presence of rhSCF, rhIL 3, rhIL 6 and rhGM CSF. Cells harvested from the culture were set in methylcellulose with cytokine cocktail of rhSCF, rhGM CSF, rhIL 3, rhIL 6, and rhEPO.Results:Cultures with the individual cytokines tested could not give rise to HPP CFC. The colonies formed in the culture system containing rhIL 3 and rhGM CSF were small in diameter and mainly consisted of macrophages.Combination of all the tested cytokines, however, resulted in large and dense colonies containing the three myeloid cell types. Furthermore, the frequency of HPP CFC in CD34 + cell culture decreased gradually with the culture time, although the absolute cell numbers increased dramatically. Conclusions:After in vitro amplification, the number of HPP CFC from CD34 + cells was maintained as evaluated by the method described. [

关 键 词：脐带血 CD34^+细胞 细胞因子 体外培养 HPP-CFC 

分 类 号：R457[医药卫生—治疗学] R331.14[医药卫生—临床医学]