- Title
- A modified MTS proliferation assay for suspended cells to avoid the interference by hydralazine and β-Mercaptoethanol
- Creator
- Wang, Yutang; Nguyen, Dinh; Anesi, Jack; Kelly, Jason; Ahmady, Fahima; Charchar, Fadi
- Date
- 2021
- Type
- Text; Journal article
- Identifier
- http://researchonline.federation.edu.au/vital/access/HandleResolver/1959.17/176588
- Identifier
- vital:15171
- Identifier
-
https://doi.org/10.1089/adt.2020.1027
- Identifier
- ISBN:1540-658X (ISSN)
- Abstract
- The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay is one of the most commonly used tests of cell proliferation. Hydralazine has been reported to interfere with the performance of the MTS assay when used on adherent cells. This study aimed to investigate whether hydralazine interferes with the performance of the MTS assay on suspended cells. THP-1 (a monocytic leukemia cell line) cells were cultured in the presence or absence of hydralazine (0, 10, 50, 100, and 500 μM) for 2 or 24 h. Cell numbers were analyzed using the MTS, trypan blue exclusion, or microscopic assays. A modified version of the standard MTS assay was established by centrifuging the cells and replacing the test medium with fresh culture medium immediately before the addition of the MTS reagent. Culture of THP-1 cells with hydralazine at concentrations of 50, 100, and 500 μM for 2 h increased absorbance (p < 0.001) in the standard MTS assay, whereas both the trypan blue exclusion assay and microscopy suggested no change in cell numbers. Culture of THP-1 cells with 100 and 500 μm hydralazine for 24 h increased absorbance (p < 0.05) in the standard MTS assay; however, trypan blue exclusion and microscopy suggested a decrease in cell numbers. In a cell-free system, hydralazine (100 and 500 μM) increased absorbance in a time- and concentration-dependent manner. The modified MTS assay produced results consistent with trypan blue exclusion and microscopy using THP-1 cells. In addition, the modified MTS assay produced reliable results when K562 and Jurkat cells were incubated with hydralazine or β-mercaptoethanol (βME). In conclusion, a simple modification of the standard MTS assay overcame the interference of hydralazine and βME when assessing suspended cells. © Copyright 2021, Mary Ann Liebert, Inc., publishers 2021. *Please note that there are multiple authors for this article therefore only the name of the Federation University Australia affiliates “Yutang Wang, Dinh Nguyen, Jack Anesi, Jason Kelly, Fahima Ahmady, Fadi Charchar" is provided in this record**
- Publisher
- Mary Ann Liebert Inc.
- Relation
- Assay and Drug Development Technologies Vol. 19, no. 3 (2021), p. 184-190. https://purl.org/au-research/grants/nhmrc/1062671
- Rights
- All metadata describing materials held in, or linked to, the repository is freely available under a CC0 licence
- Rights
- Copyright @ 2021, Mary Ann Liebert, Inc.
- Rights
- Open Access
- Subject
- 0304 Medicinal and Biomolecular Chemistry; 0306 Physical Chemistry (Incl. Structural); 1115 Pharmacology and Pharmaceutical Sciences; Cytotoxicity; Hydralazine; MTS proliferation assay; Suspension cells
- Full Text
- Reviewed
- Funder
- This work was funded by grants from the National Health and Medical Research Council of Australia (1062671). J.G. holds a Practitioner Fellowship from the National Health and Medical Research Council of Australia (NHMRC; 1117061) and a Senior Clinical Research Fellowship from the Queensland Government, Australia.
- Hits: 5450
- Visitors: 5556
- Downloads: 453
| Thumbnail | File | Description | Size | Format | |||
|---|---|---|---|---|---|---|---|
| View Details Download | SOURCE1 | Published version | 600 KB | Adobe Acrobat PDF | View Details Download |