脐血造血干/祖细胞移植SCID小鼠的实验研究  

The experimental study of engraftment in SCID mice of human umbilical cord blood hematopoietic stem/progenitor cells

作　　者：苏丽萍[1] 郝福荣[1] 张永芳[1] 缪竞诚[1] 白艳艳[1] 张占英[1] 张澜生[1] 

机构地区：[1]苏州大学生物技术系细胞免疫中心,苏州,215007

出　　处：《中国免疫学杂志》2002年第6期407-409,413,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金项目 (No.3 9970 2 44 )

摘　　要：目的 :检测扩增后脐血造血干 祖细胞的体内移植能力和造血活性 ,建立脐血细胞体外扩增优化方案和体内移植的SCID小鼠模型。方法 :采用无基质接触的液体悬浮培养方法扩增脐血CD34+ 细胞 ,将扩增前后的细胞移植给预先经过亚致死量辐照的SCID小鼠 ,4w后通过免疫荧光标记、PCR等检测存活小鼠体内的人源细胞。结果 :连续培养一定时间后 ,FL +TPO +SCF +IL 6组脐血细胞得到持续扩增 ,并能维持一定比例的CD34+ 细胞 ;SCF +IL 3+IL 6 +GM CSF +EPO组在第 2周时集落形成数已降低 ,第 4周时集落形成的细胞、CD34+ 细胞已基本检测不到。移植至少 4w后 ,在存活小鼠体内检测到人CD4 5 + 细胞和Alu基因。结论 :因子组合FL +TPO +CSF +IL 6可以有效扩增脐血CD34+ 细胞 。Objective:To detect the engraftment capability and hematopoietic potential of the expanded cells,establish an optimized project of ex vivo expansion and the SCID mice model of UCB transplantation.Methods:Human cord blood CD34 + cells were clutured in strom free liquid culture system,the sublethally irradiated SCID mice were injected with human expanded cord blood cells by tail vein.Four weeks after inocuration,BM cells of the survived animals are obtained from both femurs and tibias and assessed for the presence of human cells by immunofluorescence staining and PCR.Results:The cells and CFUs of group FL+TPO+SCF+IL 6 were increased significantly,and maintained some proportional CD34 + cells.The CFU of group SCF+IL 6+IL 3+GM CSF+EPO had reduced in week 2,CFC and CD34 + cells and Alu gene were detected in SCID mice BM.Conclusion:The cytokine combination with FL+TPO+SCF+IL 6 can effectively expand UCB CD34 + cells,and the expanded cells can engraft the BM of sublethally irradiated SCID mice and reconstitute hematopoiesis.

关 键 词：实验研究 脐血 扩增 移植 造血干细胞 造血祖细胞 

分 类 号：R331.2[医药卫生—人体生理学] R392.4[医药卫生—基础医学]