一种用于在淋巴细胞增殖和淋巴因子测定中确定细胞生长的荧光测定法。

A fluorometric assay for determining cell growth in lymphocyte proliferation and lymphokine assays.

作者信息

Dotsika E N, Sanderson C J

机构信息

National Institute for Medical Research, Mill Hill, London, U.K.

出版信息

J Immunol Methods. 1987 Dec 4;105(1):55-62. doi: 10.1016/0022-1759(87)90413-3.

DOI:10.1016/0022-1759(87)90413-3

PMID:3316407

Abstract

A microplate method for assessing cell growth and viability based on the hydrolysis of fluorogenic substrates by cell esterases has been investigated. Living cells incubated with fluorescein diacetate or 4-methylumbelliferyl heptanoate generate a fluorescent product which is proportional to the number of cells. This can be used as a simple and economical readout for various bioassays such as, for example, the assessment of IL-2 and IL-3 on factor-dependent cell lines, and on antigen- and mitogen-stimulated proliferation of lymphocytes. This fluorometric assay has similar sensitivity to the measurement of [3H]thymidine uptake and greater sensitivity than standard colorimetric assays. Incubation with MUH for a period of 30-60 min at 22 degrees C is adequate.

摘要

一种基于细胞酯酶对荧光底物的水解作用来评估细胞生长和活力的微孔板方法已被研究。用二乙酸荧光素或庚酸4-甲基伞形酮酯孵育活细胞会产生一种与细胞数量成比例的荧光产物。这可以用作各种生物测定的简单且经济的读数，例如，评估IL-2和IL-3对因子依赖性细胞系的作用，以及对抗原和丝裂原刺激的淋巴细胞增殖的作用。这种荧光测定法对[3H]胸腺嘧啶核苷摄取量的测量具有相似的灵敏度，并且比标准比色测定法具有更高的灵敏度。在22℃下与MUH孵育30 - 60分钟就足够了。