脐血造血干/祖细胞移植SCID小鼠的实验研究

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 ,而且扩增后的细胞具有较高的移植效率和造血活性

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

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.