High-Content Screening: A Decade of Evolution

Thomas, Nick

doi:10.1177/1087057109353790

Cited by 45 publications

(35 citation statements)

References 66 publications

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“…High-content imaging-based in vitro toxicity assays show promise for safety and efficacy testing as they can be performed using highthroughput systems for simultaneous screening of many compounds. 24 High-content imaging has been used with primary hepatocytes 15 and immortalized cell lines. 16,25,26 In these studies, hepatotoxicity was evaluated using morphological and biochemical readouts, including cell count, nuclear shape, mitochondria potential (MP), Ca 2 + content, and cell permeability.…”

Section: Introductionmentioning

confidence: 99%

High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells

Sirenko¹,

Hesley²,

Rusyn

et al. 2014

ASSAY and Drug Development Technologies

118

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Development of predictive in vitro assays for early toxicity evaluation is extremely important for improving the drug development process and reducing drug attrition rates during clinical development. High-content imaging-based in vitro toxicity assays are emerging as efficient tools for safety and efficacy testing to improve drug development efficiency. In this report we have used an induced pluripotent stem cell (iPSC)-derived hepatocyte cell model having a primary tissue-like phenotype, unlimited availability, and the potential to compare cells from different individuals. We examined a number of assays and phenotypic markers and developed automated screening methods for assessing multiparameter readouts of general and mechanism-specific hepatotoxicity. Endpoints assessed were cell viability, nuclear shape, average and integrated cell area, mitochondrial membrane potential, phospholipid accumulation, cytoskeleton integrity, and apoptosis. We assayed compounds with known mechanisms of toxicity and also evaluated a diverse hepatotoxicity library of 240 compounds. We conclude that high-content automated screening assays using iPSC-derived hepatocytes are feasible, provide information about mechanisms of toxicity, and can facilitate the safety assessment of drugs and chemicals.

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Section: Introductionmentioning

confidence: 99%

High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells

Sirenko¹,

Hesley²,

Rusyn

et al. 2014

ASSAY and Drug Development Technologies

118

View full text Add to dashboard Cite

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“…High Content Analysis (HCA) [21], [22], flow cytometry [23], single cell genomics [24] and other “phenotypic” methods provide the capability to measure multiple biomarkers in large numbers of individual cells. In particular, HCA can be used to profile individual cells within tissues and small animal models, as well as in 2D and 3D arrays of cells [15], [25].…”

Section: Introductionmentioning

confidence: 99%

Identifying and Quantifying Heterogeneity in High Content Analysis: Application of Heterogeneity Indices to Drug Discovery

et al. 2014

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One of the greatest challenges in biomedical research, drug discovery and diagnostics is understanding how seemingly identical cells can respond differently to perturbagens including drugs for disease treatment. Although heterogeneity has become an accepted characteristic of a population of cells, in drug discovery it is not routinely evaluated or reported. The standard practice for cell-based, high content assays has been to assume a normal distribution and to report a well-to-well average value with a standard deviation. To address this important issue we sought to define a method that could be readily implemented to identify, quantify and characterize heterogeneity in cellular and small organism assays to guide decisions during drug discovery and experimental cell/tissue profiling. Our study revealed that heterogeneity can be effectively identified and quantified with three indices that indicate diversity, non-normality and percent outliers. The indices were evaluated using the induction and inhibition of STAT3 activation in five cell lines where the systems response including sample preparation and instrument performance were well characterized and controlled. These heterogeneity indices provide a standardized method that can easily be integrated into small and large scale screening or profiling projects to guide interpretation of the biology, as well as the development of therapeutics and diagnostics. Understanding the heterogeneity in the response to perturbagens will become a critical factor in designing strategies for the development of therapeutics including targeted polypharmacology.

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“…28,31 The increased use of nonengineered cells for screening, rather than target overexpression systems, requires high-quality antibodies to establish endogenous target expression. In addition, highquality tool antibodies are required for high-content assays, 32 to provide high-throughput, multiparametric readouts of protein expression and cell-signaling events as a measure of compound effects.…”

Section: Antibody Tools In Phenotypic Screeningmentioning

confidence: 99%

The Use of Antibodies in Small-Molecule Drug Discovery

et al. 2014

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Antibodies are powerful research tools that can be used in many areas of biology to probe, measure, and perturb various biological structures. Successful drug discovery is dependent on the correct identification of a target implicated in disease, coupled with the successful selection, optimization, and development of a candidate drug. Because of their specific binding characteristics, with regard to specificity, affinity, and avidity, coupled with their amenability to protein engineering, antibodies have become a key tool in drug discovery, enabling the quantification, localization, and modulation of proteins of interest. This review summarizes the application of antibodies and other protein affinity reagents as specific research tools within the drug discovery process.

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High-Content Screening: A Decade of Evolution

Cited by 45 publications

References 66 publications

High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells

High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells

Identifying and Quantifying Heterogeneity in High Content Analysis: Application of Heterogeneity Indices to Drug Discovery

The Use of Antibodies in Small-Molecule Drug Discovery

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