造血干/祖细胞体外定向诱导分化为粒细胞的研究  

Study on Granulocytes Derived from Induction of Committed Differentiation of Hematopoietic Stem/Progenitor Cells Ex Vivo

作　　者：冯凯[1] 初飞[1] 南雪[1] 袁红丰[1] 王冬梅[1] 张锐[1] 白慈贤[1] 陈琳[1] 裴雪涛[1] 

机构地区：[1]军事医学科学院输血研究所

出　　处：《中国实验血液学杂志》2002年第6期492-495,共4页Journal of Experimental Hematology

基　　金：国家重点基础研究发展规划项目 (九七三 )基金资助项目 (编号G19990 5390 3);国家杰出青年科学基金资助项目(编号 3982 5111)

摘　　要：为探讨利用人造血干 /祖细胞体外定向诱导分化的人粒细胞应用于临床大剂量放、化疗后预防感染的可行性 ,利用源自人胎盘脐带血的CD34+ 细胞及SCF ,IL 3 ,IL 6和G CSF的细胞因子组合的培养条件 ,体外进行向粒细胞的诱导分化。结果表明 ,在该体系条件下可诱导分化出约 1 0 0 0倍数量的细胞 ,流式细胞仪检测诱导效率可达 60 %以上 ,且诱导分化的细胞染色体未出现异常 ,裸鼠致瘤实验表明该细胞不具致瘤性 ,细胞体外生长 1 4 - 2 1天为高峰 ,33天后细胞不再增殖 ,且诱导分化出的细胞具有吞噬细菌的功能。结论 :利用细胞工程的方法体外“生产”出的粒细胞具有应用安全性 ,且具备生理状态下产生的此种细胞应具备的基本功能 。To evaluated the feasibility of preventing infection after high dose chemotherapy and radiotherapy using the granulocytes derived from differentiated from hematopoietic stem/progenitor cells ex vivo, human CD34 positive cells were isolated from umbilical cord blood by using a high gradient magnetic cell sorting system(MACS), and the cells were committedly defferentiated with hematopoietic cytokines(SCF+IL 3+IL 6+G CSF) in a liquid culture system. The expanded cell number, ratio of the viable cells, chromosome and phenotype of the differentiated cells and safety analysis of expanded cells were detected by using cell count, trypan blue exclusion test, karyotype analysis, flow cytometry and tumorigenic model of nude mice, respectively. The results showed that the combination of cytokines increased cell number by (1006 4±103 2) folds and flow cytometric analysis showed myeloid marker CD11b expressed in the about 60% cells. The growth peak of differentiated cells was at 14 days of culture and decreased at about 33 days. No abnormality was found in the karyotype analysis of expanded cells. No tumor was found in the nude mice injected with expanded cells after 35 days and the expanded cells had the ability of phagocytizing bacteria. It is concluded that the cells, differentiated from CD34 + cells, expanded ex vivo possess the function of granulocyte and it was safe for clinical trial.

关 键 词：CD34^+细胞 造血干/祖细胞 体外定向 诱导分化 粒细胞 研究 

分 类 号：R331.14[医药卫生—人体生理学]