一种基于3T3-F442A细胞分化的简单灵敏的生长激素活性检测方法。

A simple and sensitive assay for GH activity based on 3T3-F442A cell differentiation.

作者信息

Juárez-Aguilar Enrique, Castro-Muñozledo Federico, Kuri-Harcuch Walid

机构信息

Department of Cell Biology, Centro de Investigación y de Estudios Avanzados del I.P.N., Apdo. Postal 14-740, 07000, México, D.F., Mexico.

出版信息

Biochem Biophys Res Commun. 2003 Nov 28;311(4):935-41. doi: 10.1016/j.bbrc.2003.10.090.

DOI:10.1016/j.bbrc.2003.10.090

PMID:14623271

Abstract

We describe a fast, sensitive, specific, and simple in vitro assay for GH biological activity, based on the differentiation of 3T3-F442A cells into adipocytes. The 3T3-F442A cells were directly plated at 1.5 x 10(4)cells/cm(2) in medium with or without various concentrations of human growth hormone (hGH). After 7 days, cells were lysed with buffer containing 0.5 % (v/v) Triton X-100, and adipose conversion was quantitated by the activity of the adipogenic enzyme glycerophosphate dehydrogenase. The assay is highly sensitive and specific for GH from different species. These culture conditions have shortened the time for the cells to undergo adipose differentiation, and they might also be useful to design and test drugs or agents that modify adipocyte differentiation or lipid metabolism, or for evaluation of cytotoxic and pharmacologic effects of drugs and other compounds.

摘要

我们描述了一种基于3T3-F442A细胞分化为脂肪细胞的快速、灵敏、特异且简单的生长激素（GH）生物活性体外检测方法。将3T3-F442A细胞以1.5×10⁴个细胞/cm²的密度直接接种于含有或不含有不同浓度人生长激素（hGH）的培养基中。7天后，用含有0.5%（v/v） Triton X-100的缓冲液裂解细胞，通过脂肪生成酶甘油磷酸脱氢酶的活性来定量脂肪转化。该检测方法对来自不同物种的GH具有高度敏感性和特异性。这些培养条件缩短了细胞进行脂肪分化的时间，它们可能也有助于设计和测试改变脂肪细胞分化或脂质代谢的药物或试剂，或用于评估药物及其他化合物的细胞毒性和药理作用。