【摘要】 本研究旨在观察几种不同细胞因子组合通过体外培养以诱导造血干/祖细胞增殖和向巨核细胞/血小板分化。应用无血清培养基(S tem Span(SFEM)体外扩增脐血CD34+细胞并向巨核细胞/血小板定向分化,将不同细胞因子组合分为3个阶段培养,并比较其培养效果。结果表明,在第一阶段的第14天时,SCF+TPO+FL+IL-3组细胞扩增倍数最高为11 000±1 000,显著高于SCF+TPO+FL组,但与SCF+TPO+IL-3组及SCF+TPO+FL+IL-3+羟皮质激素组相比较无显著差异;在第二培养阶段的第7天时,SCF+TPO+FL+IL-11组巨核细胞系扩增倍数为204666.7±11718.9,显著高于SCF+TPO+FL+IL-3组,与SCF+TPO+FL+IL1-1+BM P4+VEGF组比较并无显著差异。在第三阶段中,SCF+TPO+FL+IL-11和SCF+TPO+FL+IL-11+BM P4+VEGF2组的CD41+血小板样细胞所占比例显著高于SCF+TPO+FL+IL-3组。结论:SCF+TPO+FL+IL-3因子组合可显著提高造血干/祖细胞的扩增倍数,SCF+TPO+FL+IL-11组合有利于巨核细胞的扩增成熟及分化,为下一步的体外培养巨核细胞/血小板体系的建立奠定了一定的基础。更多还原

【Abstract】 In order to investigate the influence of cytokine combinations on proliferation and differentiation of human umbilical cord blood CD34+ cells into megakaryocytpes/platelets in vitro,the CD34+ cells from human umbilical cord blood were amplified in serum-free medium StemSpan SFEM supplemented with several cytokine combinations by three-phase culture system.The effects of the cytokine combinations were compared.The results showed that at day 14 of the first culture phase,the CD34+ cells cultured with cytokine combinations SCF+TPO+FL+IL-3 were amplified(11 000±1 000) times,which were significantly higher than that of cells cultured with SCF+TPO+FL,but were not significantly different from that of cells cultured with SCF+TPO +IL-3 or SCF+TPO+FL+IL-3+ hydroxycorticoids.At day 7 of the second culture phase,the CD34+ cells cultured with cytokine combination SCF+TPO+FL+IL-11 were amplified by(204666.7±11718.9) times,which were significantly higher than that of cells cultured with SCF+TPO+FL+IL-3,but were not significantly different from that of cells cultured with SCF+TPO+FL+IL-11+BMP4+VEGF.At day 3 and day 6,the CD34+ platelet-like cells accounted for about(39.8±1.9)%,(39.7±2.6)% and(25.5±1.4)%,(23.1±3.5)% cultured with SCF+TPO+FL+IL-11 and SCF+TPO+FL+IL-11+BMP4+VEGF,and significantly higher than that of the cells cultured with SCF+TPO+FL+IL-3.It is concluded that the cytokine combination of SCF+TPO+FL+IL-3 is most suitable cytokines combination for the amplification of CD34+ hematopoietic progenitor cells.The cytokine combination of SCF+TPO+FL+IL-11 is preferred for the proliferation and differentiation of megakaryocytes,this study lays an experimental basis for investigeting the proliferation and differentiation of CD34+ into megakaryocytes/platalets in vitro.更多还原