Dorman B P, Shimizu N, Ruddle F H
Proc Natl Acad Sci U S A. 1978 May;75(5):2363-7. doi: 10.1073/pnas.75.5.2363.
Somatic cell hybrids of human fibroblasts and mouse A9 cells, carrying only a portion of the human X chromosome in a mouse chromosome background, were injected into C3H mice. The resulting mouse anti-hybrid cell antisera contain antibodies found to be human specific and to react with only those hybrid cells carrying the human X chromosome, as confirmed by essentially perfect concordance between antibody binding assayed by indirect immunofluorescence and presence of the human X-linked enzyme hypoxanthine phosphoribosyl-transferase determined by autoradiographic assay of [3H] hypoxanthine utilization. Heterogeneous mixtures of hybrid cells may be analyzed into fluorescent (X plus) and nonfluorescent (X minus) subpopulations and fractionated viably by using a fluorescence-activated cell sorter.
将仅在小鼠染色体背景中携带一部分人类X染色体的人成纤维细胞与小鼠A9细胞的体细胞杂种注射到C3H小鼠体内。通过间接免疫荧光测定的抗体结合与通过[3H]次黄嘌呤利用的放射自显影测定确定的人类X连锁酶次黄嘌呤磷酸核糖转移酶的存在之间基本完全一致,证实所得的小鼠抗杂种细胞抗血清含有被发现具有人类特异性且仅与携带人类X染色体的那些杂种细胞反应的抗体。杂种细胞的异质混合物可以通过使用荧光激活细胞分选仪分析为荧光(X加)和非荧光(X减)亚群并进行活细胞分级分离。