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本研究提出两阶段法巨核细胞分化模型:首先将人外周血来源CD34+细胞在Cocktail或CC100两种增殖培养液中培养3、4、5或6 d,然后转入含有TPO和SCF的分化培养液中继续培养7、8或9 d。培养结束后通过比较细胞增殖、分化及成熟情况,优化培养条件。结果显示最优的诱导分化条件是,CD34+细胞在Cocktail培养液中扩增3 d后转入分化培养液中继续培养7 d;所获得的CD41+细胞和多倍体细胞数量是起始CD34+细胞的16倍和3倍;该条件下获得的CD41+和多倍体细胞数量显著高于常用的TPO法和TPO+SCF培养法。因此,运用本研究优化的两阶段培养模式能获得比单阶段直接诱导法更多的CD41+细胞和多倍体细胞,为巨核细胞和血小板相关的理论、临床研究提供新的更高效的巨核细胞体外扩增、分化模型。
In this study, a two-stage megakaryocyte differentiation model was proposed. Firstly, human peripheral blood-derived CD34 + cells were cultured for 3, 4, 5 or 6 days in Cocktail or CC100 proliferation medium and then transferred to differentiation medium containing TPO and SCF Continue training for 7, 8 or 9 days. After the end of culture, cell proliferation, differentiation and maturation were compared to optimize the culture conditions. The results showed that CD34 + cells were expanded in Cocktail broth for 3 days and then transferred to differentiation medium for 7 days. The number of CD41 + cells and polyploid cells obtained was CD34 + 16 times and 3 times respectively. The number of CD41 + and polyploid cells obtained under these conditions were significantly higher than those of TPO and TPO + SCF methods. Therefore, using the two-stage culture model optimized in this study, more CD41 + cells and polyploid cells can be obtained than the single-stage direct induction method, providing new and more efficient megakaryocytes for megakaryocyte and platelet-related theories and clinical studies in vitro Amplify and differentiate the model.