Limits...
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.


LDH Assay Z’-factor Analysis. CytoTox-ONE® Assay reagent (Promega Corporation) was added to HepG2 cells treated with 3.125µM staurosporine. Medium served as vehicle control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2802765&req=5

Figure 3: LDH Assay Z’-factor Analysis. CytoTox-ONE® Assay reagent (Promega Corporation) was added to HepG2 cells treated with 3.125µM staurosporine. Medium served as vehicle control.

Mentions: The implementation of LDH assays for cytotoxicity testing in HTS has been historically limited because most commercial assay protocols require a cell-free sample transferred into a separate assay plate. Reagents that are compatible with viable cells are available to detect release of LDH, thus enabling a single addition homogeneous assay format [45]. Now LDH release can be routinely measured in HTS formats after a short incubation (10-20 minutes) using a fluorometer equipped with a 560nm excitation source and a 590nm collection filter (Fig. 3) [46].


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)

LDH Assay Z’-factor Analysis. CytoTox-ONE® Assay reagent (Promega Corporation) was added to HepG2 cells treated with 3.125µM staurosporine. Medium served as vehicle control.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: LDH Assay Z’-factor Analysis. CytoTox-ONE® Assay reagent (Promega Corporation) was added to HepG2 cells treated with 3.125µM staurosporine. Medium served as vehicle control.
Mentions: The implementation of LDH assays for cytotoxicity testing in HTS has been historically limited because most commercial assay protocols require a cell-free sample transferred into a separate assay plate. Reagents that are compatible with viable cells are available to detect release of LDH, thus enabling a single addition homogeneous assay format [45]. Now LDH release can be routinely measured in HTS formats after a short incubation (10-20 minutes) using a fluorometer equipped with a 560nm excitation source and a 590nm collection filter (Fig. 3) [46].

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.