FL与SCF、IL-3、GM-CSF及EPO协同调控脐血造血干/祖细胞的体外扩增

Study on FL Collaborated with SCF,IL-3,GM-CSF and EPO Regulating the Expansion of Umbilical Cord Blood Hematopoietic Stem Cells in Vitro

目的 探讨造血细胞因子的不同组合对脐血造血干 /祖细胞的扩增作用。方法 应用免疫磁珠法分离纯化脐血CD3 4+ 造血干 /祖细胞 ,在体外液体培养体系中经各种不同细胞因子组合扩增 1周 ,用流式细胞仪检测CD3 4+ 造血干 /祖细胞并进行甲基纤维素法半固体培养 2周 ,在倒置显微镜下计数集落产率。结果 在FL、SCF、IL 3、GM CSF、EPO造血细胞因子的不同组合下脐血造血干 /祖细胞得以扩增 ,以IL 3+SCF +FL +EPO组的扩增效率最高 ,其有核细胞数、CD3 4+ 细胞及CFU GM、BFU E集落分别扩增 46 2± 175、3.47± 1.6 4、2 6 4± 10 5和 12 8± 6 7倍。结论 FL对扩增CD+ 3 4细胞具有较强的协同作用 ,合理的细胞因子组合扩增的脐血造血干 /祖细胞可成为异基因造血干细胞移植的主要来源。

Object To explore the effect of the different cytokine combinations on the expansion of hematopoietic stem cells of umilical cord blood(UCB).Method CD_ 34 + cells were isolated from umilical cord blood by immuno-magnetic cell sorting system(MACS) and expanded with the different combinations of cytokines in a short-term liquid culture system and in semisolid culture system for 1～2 weeks.Results In the presence of the combination of cytokine including FL,SCF,IL-3,GM-CSF and EPO,the total nucleated cell counts,CD_ 34 + cells,CFU-GM and BFU-E increased 462±175,3.47±1.64,264±105 and 128±67 times respectively.The expansion efficiency was the highest in the combination of IL-3+SCF+FL+EPO.Coulsions The data demonstrate that FL shows a strong synergic potential for expanding CD_ 34 + cells.Umbilical cord blood hematopoietic stem/progenitor cells expanded in reasonable combination of cytokines may become the main source for adult hematopoietic stem cells transplantation.