论文部分内容阅读
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
OBJECTIVE: To detect the in vivo transplanting ability and hematopoietic activity of expanded hematopoietic stem / progenitor cells from umbilical cord blood and to establish an optimized in vitro expansion program of umbilical cord blood cells and SCID mouse model in vivo. Methods: Umbilical cord blood CD34 + cells were expanded by liquid suspension culture without substrate contact. The pre-and post-expansion cells were transplanted to pre-irradiated SCID mice and survived 4 weeks later by immunofluorescence and PCR Murine human cells. Results: After continuous culture for a certain period of time, the number of cord blood cells in FL + TPO + SCF + IL 6 group continued to expand and maintain a certain proportion of CD34 + cells; in SCF + IL 3 + IL 6 + GM CSF + EPO group, When the number of colony formation has decreased, the first 4 weeks colony-forming cells, CD34 + cells have been basically undetectable. After transplantation for at least 4 w, human CD4 + cells and Alu gene were detected in surviving mice. Conclusion: The factor combination FL + TPO + CSF + IL 6 can effectively expand cord blood CD34 + cells, and the expanded cells have higher transplantation efficiency and hematopoietic activity