The effects of erythropoietin and recombinant cytokines (G-CSF, SCF, IL-3 and GM-CSF) on colony formation and self-renewal by erythroid burst- forming units (BFU-E) and granuiocyte-macrophage progenitors (CFU-GM) from mobilized peripheral blood progenitor cells (PBPCs) of the normai donors and patients were investigated. To better understand how combinations of the cytokines may be appropriate for stem cell manipulation, a model involving replating of individual BFU-E and CFU-GM colonies has been used to study the effects of cytokines on colony formation and self-renewal. Based on granulocyte colony-stimulating factor (G-CSF) and erythropoietin(EPO) alone serve as a baseline with which to compare the effects of the combinations with stem cell factor (SCF), interleukin-3 (IL-3) or granulocyte-macrophage colony-stimulating significantly increased frequency of CFU-GM in the presence of SCF, IL-3, and GM-CSF used in conjunction with G-CSF.(area under the curve of subcolony distribution) in the presence of G-CSF combined with other cytokines. GMix was identifielas the optimal combination of cytokines for both the expansion of CFU-GM as well as the expansion of clonogenic progenitor 0.0067).%本研究探讨红细胞生成素(EPO)和重组细胞因子(G-CSF,SCF,IL-3和GM-CSF)对患者和健康供者动员后的外周造血祖细胞的红系爆式祖细胞(BFU-E)和粒巨噬系祖细胞(CFU-GM)的集落形成和自我更新的作用.为了更好的了解上述那一种细胞因子和如何联合对干细胞的扩增更为有效,采用甲基纤维素半固体培养法,观察在单独用EPO、G-CSF的基础上,比较联合SCF,IL-3和GM-CSF对BFU-E和CFU-GM的作用.结果显示:①在EPO+IL-3和EPO+SCF+IL-3组,外周血BFU-E自我更新显著增加,而EPO+SCF组则明显增加.②患者与正常供者之间BFU-E的自我更新无显著差别.③G-CSF联合SCF,IL-3和GM-CSF后CFU-GM集落形成显著增加.患者组仅G-CSF本身可使CFU-GM集落形成显著增加,而G-CSF+SCF和GMix组CFU-GM集落形成明显增加.④当G-CSF联合SCF,IL-3和GM-CSF可以显著增加CFU-GM的自我更新(AUC).GMix是CFU-GM的集落形成和祖细胞扩增的较佳组合.⑤正常供者比患者有较高的AUC即自我更新,特别是在G-CSF组比患者具有显著差异(P=0.0067).
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