Developing high-content assays for hepatoxicity with iPSC-derived cells
Members from Molecular Devices (Sunnyvale, CA) and the University of North Carolina (Chapel Hill, CA), were able to use hepatocytes derived from induced pluripotent stem cells (iPSCs) to build a hepatotoxicity library of 240 compounds. In their efforts, they sought to design a high-content imaging-based in vitro toxicity assay that was compatible with iPSC-sourced cells to prove that such cells were viable models for similar assays in the future and to facilitate improvements in the drug development process, namely the safety assessment of potentially harmful substances.
In the development of the aforementioned procedure, the initial steps were to determine which assays and phenotypic markers to examine in each treatment of the hepatocytes. Observations included cell viability, cytoskeleton integrity, and mitochondrial membrane potential. With the desired parameters in mind, the researchers could design corresponding automated screening and imaging methods for each one, and then select and assemble the appropriate materials for each assay.
The reagent selected for observing mitochondrial membrane potential was JC-10, which was an ideal choice considering the live hepatocytes and the specific requirements of the high-content assay. JC-10's most convenient feature over alternative probes for mitochondrial conditions is its high aqueous solubility, which allows it to be easily compartmentalized within mitochondria, and be especially well suited for in vitro platforms.
Sirenko Oksana, Hesley Jayne, Rusyn Ivan, and Cromwell Evan F."High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell-Derived Cells". ASSAY and Drug Development Technologies. February 2014, 12(1): 43-54. doi:10.1089/adt.2013.520.
JC-10. AAT Bioquest, n.d. Web. 24 June 2016
Original created on October 4, 2016, last updated on October 4, 2016