Tsukada Y, Yoshida M C, Koura M, Isaka H, Sekiguchi T, Hirai H
Ann N Y Acad Sci. 1983;417:149-57. doi: 10.1111/j.1749-6632.1983.tb32859.x.
Cytoplasmic hybrid (cybrid) cell lines were formed by fusing whole cells of rat yolk sac tumor cell line (EST-II) with cytoplasts of mouse fibroblastic cell line (B-82cap), a variant of mouse L cell line that is deficient in thymidine kinase (TK-) and resistant to chloramphenicol (capr). The cybrid cell line with the nucleus of EST-II and cytoplasma of B-82cap was successfully obtained using double selection with HAT and chloramphenicol-containing medium. The cybrid's ability to synthesize proteins such as albumin, alpha-fetoprotein, gamma-EST, and gamma-GTP was found to be approximately one-fourth that of the nuclear donor, EST-II, at a early time of growth in culture, but this was followed by a gradual increase during the period of observation. The nude mice undergoing subcutaneous transplantation of 1 X 10(6) cells of EST-II and cybrid were killed by the tumor growth, with the survival time being 56 +/- 11 and 105 +/- 25 days, respectively. The histologic findings of cybrid tumor closely resembled those of the nuclear donor, EST-II. These facts suggest that factors from both the nucleus and cytoplasma will be able to affect the gene expression of the cybrid cell line.
细胞质杂种(cybrid)细胞系是通过将大鼠卵黄囊肿瘤细胞系(EST-II)的全细胞与小鼠成纤维细胞系(B-82cap)的细胞质体融合而成的,B-82cap是小鼠L细胞系的一个变体,缺乏胸苷激酶(TK-)且对氯霉素(capr)具有抗性。利用含有HAT和氯霉素的培养基进行双重选择,成功获得了具有EST-II细胞核和B-82cap细胞质的杂种细胞系。发现在培养早期,杂种细胞合成白蛋白、甲胎蛋白、γ-EST和γ-GTP等蛋白质的能力约为核供体EST-II的四分之一,但在观察期内逐渐增加。对接受1×10⁶个EST-II细胞和杂种细胞皮下移植的裸鼠,肿瘤生长导致其死亡,生存时间分别为56±11天和105±25天。杂种细胞肿瘤的组织学发现与核供体EST-II的非常相似。这些事实表明,细胞核和细胞质中的因子都能够影响杂种细胞系的基因表达。