Online Purchasing Account You are logged on as Guest. LoginRegister a New AccountShopping cart (Empty)
United States 

Drug toxicity screening using induced pluripotent stem cells and high-content analysis


Toxicity, and more specifically hepatotoxicity, is a major concern for the pharmaceutical industry. Both drug candidates and approved drugs face the issue of induced toxicity to the liver. Indeed, harmful side-effects to the liver is the top reason why previously approved drugs are withdrawn from the market. Over 1,000 drugs have known liver toxicity and are responsible for more than one third of acute liver failures in the United States. Hepatotoxicity is a major reason for inadequacies of drug candidates during pre-clinical and clinical stages of development. Though the current process for toxicity testing largely evolves around the use of in vivo animal work, more and more studies rely on human induced pluripotent stem cell (iPSC)-derived hepatocytes as a viable and up-and-coming alternative of determining toxicity. Animal studies take more time and money than do stem cell testing and they lack the reproducibility of results in human models. Induced pluripotent stem cell-derived hepatocytes have a primary tissue-like phenotype, constant and indefinite disposal, and the potential to establish genotype-specific cells from different individuals, which make them an interesting means of predicting drug-induced toxicity. Because these hepatocytes can be procured from individuals with a range of genotypes, it allows for direct comparisons between genotypes and pre-screening for patients to explore potential negative reactions to drugs. These cells are also stable in culture, and display reliable and reproducible performance.

Induced pluripotent stem cells are most useful when combined with a high throughput in vitro assay that can recognize hepatotoxicity. Using Enzo’s SCREEN-WELL® Hepatotoxicity library, Dr. Sirenko and colleagues from Molecular Devices developed a method called multi-parametric hepatotoxicity (MPH) by which hepatotoxicity can be measured based on several criteria at once. Cell viability, nuclear shape, average and integrated cell area, mitochondrial membrane potential, autophagy, phospholipid accumulation, cytoskeleton integrity, and apoptosis are the specifications by which liver toxicity is measured. If used in combination with high-throughput systems and toxicity assays, high-content imaging has the benefit of being able to screen many compounds at a time to assess multiple parameters including small phenotypic changes. This type of compound screening is crucial in the early stages of drug development in order to determine the potential toxic effects of the drug. The authors of this study measured assay sensitivity and specificity of the hepatotoxic compounds included in the library. Assay sensitivity represents the percentage of confirmed hepatotoxic compounds out of the 208 compounds expected to be hepatotoxic. Assay specificity is the percentage of confirmed non-hepatotoxic compounds out of the 30 compounds included as negative controls. A hit on a tested compound was defined as a result when the response parameter was greater than three standard deviations from the DMSO control, which also demonstrated an effect in more than one of the specified parameters.

Hits included classes of drugs such as anticancer drugs, neuroleptics, hormonal drugs, anti-histamines, statins, cardiac drugs, anti-fungal drugs, known toxins, antibiotics and anti-inflammatory drugs. The aforementioned multi-parametric approach was implemented to create a ranking among the tested chemicals based on relative safety as it relates to hepatotoxicity potential. The results were represented graphically using the ToxPi method to provide a visual illustration of results. From this investigation, a workflow assay protocol has been developed that allows for identification of both general and mechanism specific toxicity. Toxicity can be analyzed in more detail using this system by combining the results of a variety of cellular feedbacks to a compound of interest.

Enzo Life Sciences offers a comprehensive product portfolio for advancing your discoveries in toxicity research with a whole portfolio of toxicity research reagents and kits, including the SCREEN-WELL® Hepatotoxicity library. In addition, Enzo offers CELLESTIAL fluorescent probes ideal for high-content screening and for monitoring apoptosis, autophagy, cell viability, mitochondria membrane potential and phospholipidosis, some of which are listed below:.

Share this TechNote

Never miss a new TechNote!

Receive our TechNotes as soon as they are published.


Follow Us!

 
comments powered by Disqus

Reference:

  1. O. Sirenko, et al. High-content assays for hepatotoxicity using induced pluripotent stem cell-derived cells. Assay Drug Dev. Technol. (2014) 12:43.

Related Products

SCREEN-WELL® Cardiotoxicity library 

Essential standards for predictive toxicology screening
HTS | Print as PDF
 
BML-2850-0500 1 Library 500 µl/well Inquire for pricing
 
BML-2850-0100 1 Library 100 µl/well Inquire for pricing
Do you need bulk/larger quantities?
 

SCREEN-WELL® Hepatotoxicity library 

Essential standards for predictive toxicology screening
HTS | Print as PDF
 
BML-2851-0500 1 Library 500 µl/well Inquire for pricing
 
BML-2851-0100 1 Library 100 µl/well Inquire for pricing
Do you need bulk/larger quantities?
 

SCREEN-WELL® Hematopoietic Toxicity library 

Essential standards for predictive toxicology screening
HTS | Print as PDF
 
BML-2852-0500 1 Library 500 µl/well Inquire for pricing
 
BML-2852-0100 1 Library 100 µl/well Inquire for pricing
Do you need bulk/larger quantities?
 

GFP-CERTIFIED® Apoptosis/Necrosis detection kit 

Multiplex assay that distinguishes between  healthy, early apoptotic, late apoptotic and necrotic cells, compatible with GFP and other green fluorescent probes
Flow Cytometry, Fluorescence microscopy, Fluorescent detection | Print as PDF
 
ENZ-51002-25 25 assays 157.00 USD
 
ENZ-51002-100 100 assays 371.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® Total ROS/Superoxide detection kit 

Directly Monitor Global Levels  of Reactive Oxygen Species (ROS), and Specifically Superoxide
Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51010 1 Kit 235.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® Total ROS detection kit 

Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51011 1 Kit 172.00 USD
Do you need bulk/larger quantities?
 

ROS-ID® Superoxide detection kit 

Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51012 1 Kit 172.00 USD
Do you need bulk/larger quantities?
 

LYSO-ID® Red cytotoxicity kit (GFP-CERTIFIED®) 

A Rapid, Quantitative and HTS‐compatible Live Cell Cytotoxicity Assay 
Fluorescence microscopy, Fluorescent detection | Print as PDF
 
ENZ-51015-KP002 1 Kit 335.00 USD
Do you need bulk/larger quantities?
 

MITO-ID® Membrane potential cytotoxicity kit  

A Real-time Mitochondrial membrane Potential Assay with Superior Sensitivity
HTS, Microplate | Print as PDF
 
ENZ-51019-KP002 1 Kit 253.00 USD
Do you need bulk/larger quantities?
 

NUCLEAR-ID® Green chromatin condensation detection kit  

A highly permeable green-emitting  dye for enhanced detection of apoptosis-induced chromatin condensation
Flow Cytometry, Fluorescence microscopy, Fluorescent detection | Print as PDF
 
ENZ-51021-K200 1 Kit 284.00 USD
Do you need bulk/larger quantities?
 

EFLUXX-ID® Green multidrug resistance assay kit 

Simple No-wash Assay for  Simultaneous Monitoring of All 3 Major ABC Transporter Proteins – MDR, BCRP & MRP
Flow Cytometry, Fluorescence microscopy | Print as PDF
 
ENZ-51029-K100 1 Kit 538.00 USD
Do you need bulk/larger quantities?
 

EFLUXX-ID® Gold multidrug resistance assay kit 

Simple no-wash assays for simultaneous monitoring of all 3 major  ABC transporter proteins – MDR, BCRP & MRP
Flow Cytometry, Fluorescence microscopy | Print as PDF
 
ENZ-51030-K100 1 Kit 538.00 USD
Do you need bulk/larger quantities?
 

CYTO-ID® Autophagy detection kit 

A no-transfection, quantitative assay for monitoring autophagy in live cells
Flow Cytometry, Fluorescence microscopy, Fluorescent detection, HTS | Print as PDF
 
ENZ-51031-0050 50 tests 205.00 USD
 
ENZ-51031-K200 200 tests 420.00 USD
Do you need bulk/larger quantities?
 

PROTEOSTAT® Aggresome detection kit  

Robust, quantitative detection of aggresomes relevant to the study of neurodegenerative disease, liver disease and  toxicology
Flow Cytometry, Fluorescence microscopy, Fluorescent detection | Print as PDF
 
ENZ-51035-0025 25 tests 108.00 USD
 
ENZ-51035-K100 100 tests 294.00 USD
Do you need bulk/larger quantities?
 

SCREEN-WELL® Nephrotoxicity library 

Essential standards for predictive toxicology screening
HTS | Print as PDF
 
ENZ-LIB100-0100 1 Library 100 µl/well Inquire for pricing
Do you need bulk/larger quantities?
 

SCREEN-WELL® Myotoxicity library 

Essential standards for predictive toxicology screening
HTS | Print as PDF
 
ENZ-LIB101-0100 1 Library 100 µl/well Inquire for pricing
Do you need bulk/larger quantities?
 

Recommend this page

 
For Research Use Only. Not for use in diagnostic procedures.
Keep in touch

©2017 Enzo Life Sciences, Inc.,