Ma W, Xue S
Department of Cell Biology, Institute of Basic Medical Sciences, CAMS, Beijing.
Chin Med Sci J. 1995 Mar;10(1):1-5.
We present a novel approach for making cybrids. By introducing neo gene expression plasmids into rabbit reticulocytes, fusing the gene transferred reticulocytes with K562 cells and selecting in G418 selection medium, a cybrid strain K-RRneo was established. Whole mount TEM study demonstrated that after cybridization, there was a reorganization of the intermediate filaments which showed a tendency to differentiate towards reticulocytes. SDS-PAGE and western blot analysis verified the above observation, in which the vimentin blot pattern of the cybrids was similar to that of reticulocytes, but totally different from that of K562 cells. Using this model, we reaffirmed the hypothesis that the erythroid differentiation factor (EDF) might be responsible for erythroid differentiation as well as the initiation of denucleation.
我们提出了一种制备胞质杂种的新方法。通过将新霉素基因表达质粒导入兔网织红细胞，将导入基因的网织红细胞与K562细胞融合，并在G418选择培养基中进行筛选，建立了胞质杂种株K-RRneo。整装透射电镜研究表明，杂交后中间丝发生了重组，呈现出向网织红细胞分化的趋势。SDS-PAGE和蛋白质免疫印迹分析证实了上述观察结果，其中胞质杂种的波形蛋白印迹模式与网织红细胞相似，但与K562细胞完全不同。利用该模型，我们再次证实了红系分化因子（EDF）可能负责红系分化以及去核起始的假说。
