Deisseroth A, Burk R, Picciano D, Anderson W F, Nienhuis A, Minna J
Proc Natl Acad Sci U S A. 1975 Mar;72(3):1102-6. doi: 10.1073/pnas.72.3.1102.
Somatic cell hybrids were generated by fusion of mouse erythroleukemia cells either to mouse L cells (B82), human fibroblasts (W1-18 VA2), or human marrow fractions enriched in erythroblasts. The hybrid cells were examined for globin gene expression by benzidine staining to detect cytoplasmic hemoglobin, and by molecular hybridization of cellular RNA to globin complementary DNA (cDNA) to detect globin messenger RNA (MRNA). The fibroblast (human or mouse) times erythroleukemia cell hybrids grown in monolayer retained most of the chromosomes of each parent. Neither cytoplasmic hemoglobin nor globin mRNA was detected in dimethylsulfoxide-treated fibroblast times erythroleukemia hybrid cells, indicating extinction of hemoglobin synthesis prior to the formation of cytoplasmic mRNA. The human marrow times mouse erythroleukemia hybrid cells grown in suspension culture contained only a few human chromosomes and exhibited low levels of hemoglobin synthesis which were amplified by 2% dimethylsulfoxide. Mouse (but not human) globin mRNA was demonstrated in these hybrid cells. The results suggest that somatic cell hybrids may be useful in searching for genetic factors which regulate activity of the globin genes.
通过将小鼠红白血病细胞与小鼠L细胞(B82)、人成纤维细胞(W1 - 18 VA2)或富含成红细胞的人骨髓组分融合,产生了体细胞杂种。通过联苯胺染色检测细胞质血红蛋白来检查杂种细胞的珠蛋白基因表达,并通过细胞RNA与珠蛋白互补DNA(cDNA)的分子杂交来检测珠蛋白信使RNA(mRNA)。单层培养的成纤维细胞(人或小鼠)×红白血病细胞杂种保留了每个亲本的大部分染色体。在二甲基亚砜处理的成纤维细胞×红白血病杂种细胞中未检测到细胞质血红蛋白和珠蛋白mRNA,表明在细胞质mRNA形成之前血红蛋白合成已消失。悬浮培养的人骨髓×小鼠红白血病杂种细胞仅含有少数人类染色体,并且表现出低水平的血红蛋白合成,该合成通过2%的二甲基亚砜得以增强。在这些杂种细胞中证实存在小鼠(而非人类)珠蛋白mRNA。结果表明体细胞杂种可能有助于寻找调节珠蛋白基因活性的遗传因子。