Journal article
Moles of a Substance per Cell Is a Highly Informative Dosing Metric in Cell Culture
PloS one, Vol.10(7), pp.e0132572-e0132572
2015
DOI: 10.1371/journal.pone.0132572
PMCID: PMC4501792
PMID: 26172833
Abstract
The biological consequences upon exposure of cells in culture to a dose of xenobiotic are not only dependent on biological variables, but also the physical aspects of experiments e.g. cell number and media volume. Dependence on physical aspects is often overlooked due to the unrecognized ambiguity in the dominant metric used to express exposure, i.e. initial concentration of xenobiotic delivered to the culture medium over the cells. We hypothesize that for many xenobiotics, specifying dose as moles per cell will reduce this ambiguity. Dose as moles per cell can also provide additional information not easily obtainable with traditional dosing metrics.
Here, 1,4-benzoquinone and oligomycin A are used as model compounds to investigate moles per cell as an informative dosing metric. Mechanistic insight into reactions with intracellular molecules, differences between sequential and bolus addition of xenobiotic and the influence of cell volume and protein content on toxicity are also investigated.
When the dose of 1,4-benzoquinone or oligomycin A was specified as moles per cell, toxicity was independent of the physical conditions used (number of cells, volume of medium). When using moles per cell as a dose-metric, direct quantitative comparisons can be made between biochemical or biological endpoints and the dose of xenobiotic applied. For example, the toxicity of 1,4-benzoquinone correlated inversely with intracellular volume for all five cell lines exposed (C6, MDA-MB231, A549, MIA PaCa-2, and HepG2).
Moles per cell is a useful and informative dosing metric in cell culture. This dosing metric is a scalable parameter that: can reduce ambiguity between experiments having different physical conditions; provides additional mechanistic information; allows direct comparison between different cells; affords a more uniform platform for experimental design; addresses the important issue of repeatability of experimental results, and could increase the translatability of information gained from in vitro experiments.
Details
- Title: Subtitle
- Moles of a Substance per Cell Is a Highly Informative Dosing Metric in Cell Culture
- Creators
- Claire M Doskey - Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, Iowa City, Iowa, 52242, United States of AmericaThomas J van 't Erve - Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, Iowa City, Iowa, 52242, United States of AmericaBrett A Wagner - Free Radical and Radiation Biology Program, Department of Radiation Oncology, The University of Iowa, Iowa City, Iowa, 52242, United States of AmericaGarry R Buettner - Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, Iowa City, Iowa, 52242, United States of America; Free Radical and Radiation Biology Program, Department of Radiation Oncology, The University of Iowa, Iowa City, Iowa, 52242, United States of America
- Resource Type
- Journal article
- Publication Details
- PloS one, Vol.10(7), pp.e0132572-e0132572
- DOI
- 10.1371/journal.pone.0132572
- PMID
- 26172833
- PMCID
- PMC4501792
- NLM abbreviation
- PLoS One
- ISSN
- 1932-6203
- eISSN
- 1932-6203
- Publisher
- Public Library of Science; United States
- Grant note
- P30 ES005605 / NIEHS NIH HHS R01 GM073929 / NIGMS NIH HHS P42 ES013661 / NIEHS NIH HHS NIH P30 CA086862 / NCI NIH HHS R01GM073929 / NIGMS NIH HHS P30 CA086862 / NCI NIH HHS R01 CA169046 / NCI NIH HHS
- Language
- English
- Date published
- 2015
- Academic Unit
- Radiation Oncology; Iowa Superfund Research Program
- Record Identifier
- 9984047777402771
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