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Rapid colorimetric assay for cell growth and survival
Abstract
A convenient way to estimate the number of viable cells growing in microtitre tray wells is to use a colorimetric assay and an automatic microplate scanning spectrophotometer. One such assay, developed by Mosmann, depends on the reduction by living cells of tetrazolium salt, MTT, to form a blue formazan product. However the original technique has several technical limitations, namely a less than optimal sensitivity, a variable background due to protein precipitation on adding an organic solvent to dissolve the blue formazan product, and a low solubility of the product. These problems have been overcome by the following modifications: avoidance of serum in the incubation medium, thus overcoming precipitation problems in the organic solvent; avoidance of phenol red in the incubation medium, thus avoiding the use of acid in the final solvent which altered the spectral properties of the formazan; elimination of the medium containing MTT after the reaction and subsequent use of pure propanol or ethanol to rapidly solubilize the formazan; use of a higher concentration of MTT; use of half-area microtitre trays to increase the spectrophotometer readings from a given amount of formazan; use of a more judicious reference wavelength in a dual wavelength spectrophotometer. With these modifications the reliability and sensitivity of the test have been increased to the point where it can in many cases replace the [3H]thymidine uptake assay to measure cell proliferation or survival in growt…
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The results of the present study are in agreement with previous studies; treatment of paracetamol caused depletion of GSH, cell damage (MDA levels) and mitochondrial dysfunction.
“The results of present study are in agreement with previous studies; we found that treatment of paracetamol caused depletion of GSH, cell damage (MDA levels) and mitochondrial dysfunction. 4,7,38”
From Caralluma umbellata Haw. protects liver against paracetamol toxicity and inhibits CYP2E1 (2017) · cites this paper as [38]
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Metadata
- Authors
- Denizot, F., & Lang, R.
- Affiliations
- Walter and Eliza Hall Institute of Medical Research (AU)
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 1986-05
- Volume
- 89
- Issue
- 2
- Pages
- 271-277
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W1629179716
- Cited by
- 4,935
- References
- 0
- Citations by year
- 2026: 119 · 2025: 152 · 2024: 167 · 2023: 166 · 2022: 173 · 2021: 183 · 2020: 191 · 2019: 166 · 2018: 193 · 2017: 178 · 2016: 180 · 2015: 189 · 2014: 205 · 2013: 174 · 2012: 189
- FWCI
- 39.32
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (R01AI017310)
- Topics
- Health Sciences › Veterinary › Small Animals › Animal testing and alternatives
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Keywords
- MTT assay, cell proliferation, cell survival, cytotoxicity assay, interleukin-2, serum-free medium
- MeSH terms
- Animals, Cell Division, Cell Line, Cell Survival, Colorimetry, Culture Media, Interleukin-2, Lymphocyte Activation, Phenolsulfonphthalein, Tetrazolium Salts, Thymidine, Mice
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