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为了解胚胎时期巨核细胞增殖分化特有的内在机制,本研究观察了在体外液体培养体系中,胎肝源CD34~+造血千/祖细胞在血小板生成素(TPO)作用下增殖分化特征与相关周期蛋白B1、D1和D3表达及细胞内水平变化的关系。结果表明:①经12天培养后,TPO使胎肝源CD34细胞数从1×10~5个/ml增加到13.12±4.06×~5个/ml,CD41~+细胞增加了95%,CD34~+细胞下降了3%,大部分细胞的DNA倍性为2N,少数为4N,无>4N巨核细胞:②在整个培养期间,周期蛋白B1表达逐渐增加,并保持在一个高水平上,培养后期,高水平的周期蛋白B1出现在GI期细胞上:③周期蛋白D1和D3表达先增加,培养后期细胞内水平下降,且以G2期细胞为主。本研究结果提示:①FPO通过上调周期蛋白B1和在所有细胞周期时限上调周期蛋白D1和D3表达,促进巨核细胞祖细胞的增殖分化;②周期蛋白B1在G2+M期的持续高水平和周期蛋白D1和D3在G2+M期的水平下降可能导致了胎肝源巨核细胞核内有丝分裂延迟或阻滞。
In order to understand the intrinsic mechanism of megakaryocyte proliferation and differentiation during embryogenesis, we observed the proliferation and differentiation characteristics of fetal liver derived CD34 ~ + hematopoietic stem / progenitor cells under thrombopoietin (TPO) and related cycles Relationship between expression of protein B1, D1 and D3 and intracellular level changes. The results showed that: (1) After 12 days incubation, TPO increased the number of CD34 + cells from 1 × 10-5 / ml to 13.12 ± 4.06 × -5 cells / ml, and CD14 + cells increased 95% + Cells decreased by 3%, most of the cells had DNA ploidy of 2N, a few of 4N, no> 4N megakaryocytes: ② During the whole culture period, the expression of cyclin B1 gradually increased and remained at a high level, , The high level of cyclin B1 appeared in the GI phase cells: ③ cyclin D1 and D3 expression increased first, the late intracellular level decreased, and the G2 phase cells. The results of this study suggest that: (1) FPO promotes the proliferation and differentiation of megakaryocyte progenitor cells by up-regulating cyclin B1 and up-regulating the expression of cyclin D1 and D3 in all cell cycle stages; (2) sustained high level of cyclin B1 in G2 + M phase and cyclin Decreased levels of D1 and D3 at G2 + M phase may result in delayed or blocked mitotic arrest in fetal liver megakaryocytes.