原红细胞在体外EPO诱导下的增殖分化特征

PROLIFERATION AND DIFFERENTIATION CHARACTERISTICS OF MURINE PROERYTHROBLASTS INDUCED IN VITRO BY EPO

目的 系统了解小鼠红细胞终末分化过程中细胞增殖和分化的动力学。方法 选用FVA细胞模型 ,利用密度梯度离心的方法从感染Friend致贫血病毒 (anemia inducingstrainofFriendvirus ,FVA)的小鼠脾脏中分离出同步的原始阶段红细胞 (FVA细胞 )。通过研究EPO诱导下细胞的生长曲线 ,3H TdR掺入率以及细胞分化过程中 β 珠蛋白 ( β globin)基因的表达和血红蛋白合成的时空关系 ,分析FVA细胞在体外EPO诱导分化过程中增殖及分化的特性。结果 FVA细胞在EPO诱导早期生长旺盛 ,诱导 2 4小时细胞数量达高峰 ,为 0时的 4倍 ;3H TdR掺入在诱导后 12小时达高峰 ;从诱导后12小时开始有β globin基因的转录 ,在 2 4小时达高峰 ,随后减低 ;从 18小时开始出现血红蛋白 ,至 48小时 ,几乎全部细胞都表达大量血红蛋白。结论 本研究结果阐明了FVA细胞在体外EPO诱导下增殖及分化的动力学 。

Objective To investigate the kinetics of proliferation and differentiation of murine erythroblasts in the terminal differentiation. Method FVA cells were used as a model. The homogenous proerythroblasts (FVA cells) were isolated by using density gradient centrifugation from the spleens of mice infected with anemia inducing strain of Friend virus (FVA). The cell growth curve, the 3H-TdR incorporation rate, and the timing relations between the expression of (β-globin) gene and the synthesis of hemoglobin were studied in detail to understand the chavataristics of the proliferation and differentiation of FVA cells induced in vitro by EPO. Result FVA cells grew rapidly at the early stage of EPO treatment, and the a cell number peaked at 24 h of culture with EPO, which was 4 that times at 0 h. The peak of 3H-TdR incorporation appeared at 12 h. The transcription of β-globin gene began at 12 h after EPO treatment, reached its highest expression level at 24 h, and then declined slightly. The hemoglobin began to appear at 18 h, and almost all cells expressed hemoglobin at 48 h after EPO treatment. Conclusion The results of this study demonstrate in detail the kinetics of proliferation and differentiation of FVA cells induced in vitro by EPO. The data provide basic information for studying on the mechanisms of erythroblasts terminal differentiation using FVA model.