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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.


Fluorescent/Fluorescent and Fluorescent Luminescent Viability and Cytotoxicity Assays. MultiTox-Fluor or MultiTox-Glo Reagents (Promega Corporation) were added to a 1536 well plate after Jurkat cells had been treated with 30µg/ml digitonin or vehicle. Each well is represented as a ratio of the viable and non-viable values measured using the chemistries.
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Figure 6: Fluorescent/Fluorescent and Fluorescent Luminescent Viability and Cytotoxicity Assays. MultiTox-Fluor or MultiTox-Glo Reagents (Promega Corporation) were added to a 1536 well plate after Jurkat cells had been treated with 30µg/ml digitonin or vehicle. Each well is represented as a ratio of the viable and non-viable values measured using the chemistries.

Mentions: A new, more amenable method for HTS environments combines the protease substrates for viability and cytotoxicity allowing for simultaneous measurement of viable and non-viable cell populations (Fig. 6). This method delivers broad linear responses (0-50,000 cells/well) and can measure as little as a 2% change in viability in both the AFC and R110 channels [59]. Furthermore, the reagent can be concentrated and delivered into assay wells in a substantially reduced volume to accommodate a third spectrally distinct reagent.


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)

Fluorescent/Fluorescent and Fluorescent Luminescent Viability and Cytotoxicity Assays. MultiTox-Fluor or MultiTox-Glo Reagents (Promega Corporation) were added to a 1536 well plate after Jurkat cells had been treated with 30µg/ml digitonin or vehicle. Each well is represented as a ratio of the viable and non-viable values measured using the chemistries.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 6: Fluorescent/Fluorescent and Fluorescent Luminescent Viability and Cytotoxicity Assays. MultiTox-Fluor or MultiTox-Glo Reagents (Promega Corporation) were added to a 1536 well plate after Jurkat cells had been treated with 30µg/ml digitonin or vehicle. Each well is represented as a ratio of the viable and non-viable values measured using the chemistries.
Mentions: A new, more amenable method for HTS environments combines the protease substrates for viability and cytotoxicity allowing for simultaneous measurement of viable and non-viable cell populations (Fig. 6). This method delivers broad linear responses (0-50,000 cells/well) and can measure as little as a 2% change in viability in both the AFC and R110 channels [59]. Furthermore, the reagent can be concentrated and delivered into assay wells in a substantially reduced volume to accommodate a third spectrally distinct reagent.

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.