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Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence

OBJECTIVE: Short-term viability assays of cultured cells in 96-well plates are routinely used to determine the cytotoxicity or safety of drugs. These are often based on the formation of chromogen, generated selectively in viable cells. The innate problems of such short-term cell viability assays inc...

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Autores principales: Garg, Sukant, Huifu, He, Kaul, Sunil C., Wadhwa, Renu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6013999/
https://www.ncbi.nlm.nih.gov/pubmed/29929534
http://dx.doi.org/10.1186/s13104-018-3512-5
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author Garg, Sukant
Huifu, He
Kaul, Sunil C.
Wadhwa, Renu
author_facet Garg, Sukant
Huifu, He
Kaul, Sunil C.
Wadhwa, Renu
author_sort Garg, Sukant
collection PubMed
description OBJECTIVE: Short-term viability assays of cultured cells in 96-well plates are routinely used to determine the cytotoxicity or safety of drugs. These are often based on the formation of chromogen, generated selectively in viable cells. The innate problems of such short-term cell viability assays include (i) effect of drugs is determined by cell density (ii) some drugs have slow/gradual effect and hence may escape such assays, (iii) cell morphology that reveal significant hints to molecular signaling underlining the effect of drugs cannot be effectively captured, (iv) long-term effect on viability and clonogenic potential of cells cannot be determined and (v) herbal extracts often possess intrinsic color that interferes with spectrophotometer estimation. In light of the ease and importance of cell culture-based assessment of drug safety and cytotoxicity, we attempted to combine the conventional cell-based assays in a way that allows multiple readouts (quantitative and qualitative) from a single experiment, and avoids the drawbacks of color interference. RESULTS: We have established and validated (using 16 types of cultured mammalian cells) a Quantitative and Qualitative Cell Viability assay in 12-well cell culture plates. It overcomes several shortcomings as discussed above and allows long-term observations on cell morphology and clonogenicity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-018-3512-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-60139992018-07-05 Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence Garg, Sukant Huifu, He Kaul, Sunil C. Wadhwa, Renu BMC Res Notes Research Note OBJECTIVE: Short-term viability assays of cultured cells in 96-well plates are routinely used to determine the cytotoxicity or safety of drugs. These are often based on the formation of chromogen, generated selectively in viable cells. The innate problems of such short-term cell viability assays include (i) effect of drugs is determined by cell density (ii) some drugs have slow/gradual effect and hence may escape such assays, (iii) cell morphology that reveal significant hints to molecular signaling underlining the effect of drugs cannot be effectively captured, (iv) long-term effect on viability and clonogenic potential of cells cannot be determined and (v) herbal extracts often possess intrinsic color that interferes with spectrophotometer estimation. In light of the ease and importance of cell culture-based assessment of drug safety and cytotoxicity, we attempted to combine the conventional cell-based assays in a way that allows multiple readouts (quantitative and qualitative) from a single experiment, and avoids the drawbacks of color interference. RESULTS: We have established and validated (using 16 types of cultured mammalian cells) a Quantitative and Qualitative Cell Viability assay in 12-well cell culture plates. It overcomes several shortcomings as discussed above and allows long-term observations on cell morphology and clonogenicity. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13104-018-3512-5) contains supplementary material, which is available to authorized users. BioMed Central 2018-06-22 /pmc/articles/PMC6013999/ /pubmed/29929534 http://dx.doi.org/10.1186/s13104-018-3512-5 Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Note
Garg, Sukant
Huifu, He
Kaul, Sunil C.
Wadhwa, Renu
Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title_full Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title_fullStr Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title_full_unstemmed Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title_short Integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
title_sort integration of conventional cell viability assays for reliable and reproducible read-outs: experimental evidence
topic Research Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6013999/
https://www.ncbi.nlm.nih.gov/pubmed/29929534
http://dx.doi.org/10.1186/s13104-018-3512-5
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