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用可诱发小鼠贫血的病毒(anemia-inducingstrainoffriend’svirus,FVA)感染小鼠,取其脾细胞,在加红细胞生成素(EPO)的培养基中培养,获得大量相对同步化的早幼、中幼、晚幼和网织红细胞,并对这些不同发生阶段的红系细胞进行红细胞分化因子(EDF)粗提液的分离和提取,以检测造血过程中,不同阶段红细胞EDF的合成。并以小鼠红白血病细胞(MEL)作为靶细胞,分析不同发育阶段红细胞EDF粗提液对MEL细胞的生长和分化的生物学效应。结果表明,不同发育阶段的红细胞中EDF含量有规律性的差别,早幼红细胞期可检测到明显的EDF活性,中幼红细胞达到高峰,从晚幼红细胞期开始下降,网织红细胞期活性明显减低。用0.1g/LEDF粗提液可抑制MEL细胞的生长和3H-TdR的掺入并出现类似红系细胞排核前核固缩等分化特征。本研究结果与我们先前提出的EDF在造血细胞分化过程中起调控作用的观点提供了又一个证据。
Mice were infected with anemia-inducing strain of virus (FVA) and their splenocytes were harvested and cultured in medium supplemented with erythropoietin (EPO) to obtain a large number of relatively synchronized premature young, middle Juvenile, late juvenile and reticulocytes were isolated and extracted from erythrocyte differentiation factor (EDF) crude extracts at different stages of erythrocytes to detect the erythrocyte EDF synthesis in different stages of hematopoietic process. The mouse erythroleukemia cells (MEL) were used as target cells to analyze the biological effects of EDF crude extracts on the growth and differentiation of MEL cells at different developmental stages. The results showed that the EDF content of erythrocytes in different developmental stages had regular differences, EDF activity was detected in early erythrocyte stage, middle and later erythrocytes peaked, and decreased from late erythrocyte stage, the reticulocyte activity was significantly decreased . With 0.1g / LEDF crude extract can inhibit the growth of MEL cells and 3H-TdR incorporation and similar erythroid cell nuclear pre-nuclear nuclear shrinkage and other differentiation characteristics. The results of this study provide yet another evidence of the regulatory role of EDF in our hematopoietic cell differentiation as previously proposed.