Zhang Ke-Ying, Liu Jiang, Jia Yan-Jun, Li Wei, Duan Lan, Gao Song-Ming, Cui Shuang, Gong Zhi-Yin, Ni Lei, Zhang Zhi-Xin
Beijing Red Cross Blood Service Center, Beijing, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Aug;19(4):1053-7.
In order to investigate the influence of cytokine combinations on proliferation and differentiation of human umbilical cord blood CD34(+) cells into megakaryocytes/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+ hydroxyl-corticosteroids. 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 investigating the proliferation and differentiation of CD34(+) into megakaryocytes/platelets in vitro.
为了研究细胞因子组合对人脐血CD34(+)细胞体外增殖及向巨核细胞/血小板分化的影响，采用三相培养体系，在添加多种细胞因子组合的无血清培养基StemSpan(SFEM)中扩增人脐血CD34(+)细胞。比较了不同细胞因子组合的作用效果。结果显示，在第一培养阶段第14天，用细胞因子组合SCF + TPO + FL + IL-3培养的CD34(+)细胞扩增了(11 000 ± 1 000)倍，显著高于用SCF + TPO + FL培养的细胞，但与用SCF + TPO + IL-3或SCF + TPO + FL + IL-3 + 羟基皮质类固醇培养的细胞无显著差异。在第二培养阶段第7天，用细胞因子组合SCF + TPO + FL + IL-11培养的CD34(+)细胞扩增了(204666.7 ± 11718.9)倍，显著高于用SCF + TPO + FL + IL-3培养的细胞，但与用SCF + TPO + FL + IL-11 + BMP4 + VEGF培养的细胞无显著差异。在第3天和第6天，用SCF + TPO + FL + IL-11和SCF + TPO + FL + IL-11 + BMP4 + VEGF培养的CD34(+)类血小板细胞分别占(39.8 ± 1.9)%、(39.7 ± 2.6)%和(25.5 ± 1.4)%、(23.1 ± 3.5)%，显著高于用SCF + TPO + FL + IL-3培养的细胞。结论：SCF + TPO + FL + IL-3细胞因子组合是扩增CD34(+)造血祖细胞最合适的细胞因子组合。SCF + TPO + FL + IL-11细胞因子组合更有利于巨核细胞的增殖和分化，本研究为体外研究CD34(+)向巨核细胞/血小板的增殖和分化奠定了实验基础
