一种改良的MTT检测法。

An improved MTT assay.

作者信息

Sladowski D, Steer S J, Clothier R H, Balls M

机构信息

FRAME Alternatives Laboratory, Department of Human Morphology, University of Nottingham Medical School, Queen's Medical Centre, UK.

出版信息

J Immunol Methods. 1993 Jan 4;157(1-2):203-7. doi: 10.1016/0022-1759(93)90088-o.

DOI:10.1016/0022-1759(93)90088-o

PMID:8423364

Abstract

The MTT assay for cell viability and cell proliferation has been modified to improve its reproducibility and accuracy. The modified test is performed on MultiScreen filtration plates, which permits the removal of the culture medium prior to formazan solubilisation, without loss of cells or formazan crystals. A 1:1 mix of DMSO and ethanol is used as the solvent, since this has the same optical refraction index as the filters, making it possible to measure the optical densities directly on the MultiScreen plate. Data obtained in the assay method using the MultiScreen plate were compared with data from studies employing the normal flat-bottomed plate, by using cells which grow in suspension. Two cell lines were used in the study. CTLL-2, which are IL-2 dependent cells of murine T cell origin, and Jurkat E.6.1 which are IL-2 producing cells of human lymphoma origin. CTLL-2 cells and the modified MTT assay were also used for evaluating the effects of different IL-2 concentrations on cell proliferation.

摘要

用于细胞活力和细胞增殖的MTT试验已被改进，以提高其重现性和准确性。改进后的试验在MultiScreen过滤板上进行，该过滤板允许在甲臜溶解之前去除培养基，而不会损失细胞或甲臜晶体。使用二甲基亚砜（DMSO）和乙醇的1:1混合物作为溶剂，因为它与过滤器具有相同的光学折射率，使得可以直接在MultiScreen板上测量光密度。通过使用悬浮生长的细胞，将使用MultiScreen板的测定方法获得的数据与采用普通平底培养板的研究数据进行比较。该研究使用了两种细胞系。CTLL-2，它是源自小鼠T细胞的白细胞介素-2（IL-2）依赖细胞，以及Jurkat E.6.1，它是源自人类淋巴瘤的产生IL-2的细胞。CTLL-2细胞和改进后的MTT试验也用于评估不同IL-2浓度对细胞增殖的影响。