细胞因子体外诱导脐血CD34^+细胞增殖并向巨核细胞/血小板分化的研究  

Influence of Cytokine Combinations on Proliferation and Differentiation of Umbilical Cord Blood CD34^+ Cells into Megakaryocytes/Platelets In Vitro

作　　者：张可莹[1] 刘江[1] 贾延军[1] 李伟[1] 段澜[1] 高颂明[1] 崔爽[1] 龚治尹[1] 倪雷[1] 张志欣[1] 

机构地区：[1]北京市红十字血液中心,北京100088

出　　处：《中国实验血液学杂志》2011年第4期1053-1057,共5页Journal of Experimental Hematology

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。In order to investigate the influence of cytokine combinations on proliferation and differentiation of human umbilical cord blood CD34＋ cells into rnegakaryocytpes/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 1300 ~ 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 CTY34 ＋ 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-I1 is preferred for the proliferation and differentiation of megakaryocytes, this study lays an experimental basis for investigefing the proliferation and differentiation of CD34 ＋ into megakaryocytes/platalets in vitro.

关 键 词：细胞因子 体外培养 血小板 脐血CD34’细胞 血小板增殖 血小板分化 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R331.143[医药卫生—基础医学]