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In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening.

Niles AL, Moravec RA, Riss TL - Curr Chem Genomics (2009)

Bottom Line: In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities.The early and routine implementation of this testing is testament to its prognostic importance for humans.We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method.

View Article: PubMed Central - PubMed

Affiliation: Research Department, Promega Corporation, 2800 Woods Hollow Road, Madison, WI, USA. andrew.niles@promega.com

ABSTRACT
In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities. The early and routine implementation of this testing is testament to its prognostic importance for humans. Although a plethora of assay chemistries and methods exist for 96-well formats, few are practical and sufficiently sensitive enough for application in high throughput screening (HTS). Here we briefly describe a handful of the currently most robust and validated HTS assays for accurate and efficient assessment of cytotoxic risk. We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method. Lastly, we discuss the advantages of combining specific HTS compatible assays into multi-parametric, same-well formats.

No MeSH data available.


Single- and Multi-parameter Viability and Cytotoxicity Assays. CellTiter-Blue®, CytoTox-ONE™, CellTiter-Glo®, MultiTox-Fluor, MultiTox-Glo and CytoTox-Glo™ Reagents (Promega Corporation) were added to Jurkat cells treated for 6 hr with serial dilutions of ionomycin and signals measured as described in technical literature.
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Figure 5: Single- and Multi-parameter Viability and Cytotoxicity Assays. CellTiter-Blue®, CytoTox-ONE™, CellTiter-Glo®, MultiTox-Fluor, MultiTox-Glo and CytoTox-Glo™ Reagents (Promega Corporation) were added to Jurkat cells treated for 6 hr with serial dilutions of ionomycin and signals measured as described in technical literature.

Mentions: Several informative combinations of viability and cytotoxicity chemistries are currently employed in HTS formats to mitigate false determinations. These multiplexes address systematic error caused either by disparate cell numbers delivered to assay wells or random compound interferences with reporter molecules [55]. Except in cases of cell-cycle arrest prior to loss of membrane integrity, or long term exposure models, measures of viability and cytotoxicity are inversely proportional (Fig. 5). Therefore, non-conforming data sets can easily “flag” problem compounds for further analysis by orthogonal methods [57].


In vitro viability and cytotoxicity testing and same-well multi-parametric combinations for high throughput screening.

Niles AL, Moravec RA, Riss TL - Curr Chem Genomics (2009)

Single- and Multi-parameter Viability and Cytotoxicity Assays. CellTiter-Blue®, CytoTox-ONE™, CellTiter-Glo®, MultiTox-Fluor, MultiTox-Glo and CytoTox-Glo™ Reagents (Promega Corporation) were added to Jurkat cells treated for 6 hr with serial dilutions of ionomycin and signals measured as described in technical literature.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2802765&req=5

Figure 5: Single- and Multi-parameter Viability and Cytotoxicity Assays. CellTiter-Blue®, CytoTox-ONE™, CellTiter-Glo®, MultiTox-Fluor, MultiTox-Glo and CytoTox-Glo™ Reagents (Promega Corporation) were added to Jurkat cells treated for 6 hr with serial dilutions of ionomycin and signals measured as described in technical literature.
Mentions: Several informative combinations of viability and cytotoxicity chemistries are currently employed in HTS formats to mitigate false determinations. These multiplexes address systematic error caused either by disparate cell numbers delivered to assay wells or random compound interferences with reporter molecules [55]. Except in cases of cell-cycle arrest prior to loss of membrane integrity, or long term exposure models, measures of viability and cytotoxicity are inversely proportional (Fig. 5). Therefore, non-conforming data sets can easily “flag” problem compounds for further analysis by orthogonal methods [57].

Bottom Line: In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities.The early and routine implementation of this testing is testament to its prognostic importance for humans.We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method.

View Article: PubMed Central - PubMed

Affiliation: Research Department, Promega Corporation, 2800 Woods Hollow Road, Madison, WI, USA. andrew.niles@promega.com

ABSTRACT
In vitro cytotoxicity testing has become an integral aspect of drug discovery because it is a convenient, costeffective, and predictive means of characterizing the toxic potential of new chemical entities. The early and routine implementation of this testing is testament to its prognostic importance for humans. Although a plethora of assay chemistries and methods exist for 96-well formats, few are practical and sufficiently sensitive enough for application in high throughput screening (HTS). Here we briefly describe a handful of the currently most robust and validated HTS assays for accurate and efficient assessment of cytotoxic risk. We also provide guidance for successful HTS implementation and discuss unique merits and detractions inherent in each method. Lastly, we discuss the advantages of combining specific HTS compatible assays into multi-parametric, same-well formats.

No MeSH data available.