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非洲爪蟾输卵管细胞原代培养的建立与特性分析

Establishment and characterization of Xenopus oviduct cells in primary culture.

作者信息

Marsh J, Tata J R

机构信息

Laboratory of Developmental Biochemistry, National Institute for Medical Research, London, United Kingdom.

出版信息

Exp Cell Res. 1987 Nov;173(1):117-28. doi: 10.1016/0014-4827(87)90337-5.

Abstract

Based on a previously established procedure for Xenopus hepatocytes, we describe tubular oviduct cells in primary culture which continue to secrete substantial quantities of egg jelly for several days, as can be visualized microscopically. Freshly isolated cells exhibited a culture shock response [A. P. Wolffe, J. F. Glover, and J. R. Tata (1984) Exp. Cell Res. 154, 581], from which they recovered by the third day in culture. This recovery was characterized by (a) the diminished synthesis of heat shock proteins hsp 70 and hsp 85, (b) the cessation of the drop in number of estrogen receptor, and (c) the enhanced rate of synthesis of cellular and secreted proteins. The oviduct estrogen receptor had the same characteristics as those in other estrogen target tissues and was present in the same amount as in adult female Xenopus hepatocytes [A. J. Perlman, A. P. Wolffe, J. Champion, and J. R. Tata (1984) Mol. Cell. Endocrinol. 38, 51]. Unlike the latter in primary culture [M. P. R. Tenniswood, P. F. Searle, A. P. Wolffe, and J. R. Tata (1983) Mol. Cell. Endocrinol. 30, 329], oviduct cell cultures did not actively metabolize either estradiol or progesterone (t1/2 approximately equal to 55 and 7 h, respectively). The successful establishment and characterization of primary cultures of both liver and oviduct cells now fulfil the conditions required for investigating the basis for tissue specificity of regulation by estrogen of Xenopus egg protein gene expression in primary cell culture.

摘要

基于之前建立的非洲爪蟾肝细胞程序,我们描述了原代培养的输卵管细胞,这些细胞在数天内持续分泌大量卵胶,这在显微镜下可见。刚分离的细胞表现出培养应激反应[A. P. 沃尔夫、J. F. 格洛弗和J. R. 塔塔(1984年),《实验细胞研究》154卷,581页],到培养第三天时它们从中恢复。这种恢复的特征是:(a)热休克蛋白hsp 70和hsp 85的合成减少;(b)雌激素受体数量下降停止;(c)细胞和分泌蛋白的合成速率提高。输卵管雌激素受体具有与其他雌激素靶组织中相同的特征,其含量与成年雌性非洲爪蟾肝细胞中的相同[A. J. 珀尔曼、A. P. 沃尔夫、J. 钱皮恩和J. R. 塔塔(1984年),《分子与细胞内分泌学》38卷,51页]。与原代培养的肝细胞不同[M. P. R. 坦尼斯伍德、P. F. 塞尔、A. P. 沃尔夫和J. R. 塔塔(1983年),《分子与细胞内分泌学》30卷,329页],输卵管细胞培养物不活跃代谢雌二醇或孕酮(半衰期分别约为55小时和7小时)。肝脏和输卵管细胞原代培养的成功建立和特性描述现在满足了在原代细胞培养中研究雌激素对非洲爪蟾卵蛋白基因表达调控的组织特异性基础所需的条件。

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