Development of high content imaging methods for cell death detection in human pluripotent stem cell-derived cardiomyocytes.

Abstract

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CM) are being investigated as a new source of cardiac cells for drug safety assessment. We developed a novel scalable high content microscopy-based method for the detection of cell death in hPSC-CM that can serve for future predictive in vitro cardio-toxicological screens. Using rat neonatal ventricular cardiomyocytes (RVNC) or hPSC-CM, assays for nuclear remodelling, mitochondrial status, apoptosis and necrosis were designed using a combination of fluorescent dyes and antibodies on an automated microscopy platform. This allowed the observation of a chelerythrine-induced concentration-dependent apoptosis to necrosis switch and time-dependent progression of early apoptotic cells towards a necrotic-like phenotype. Susceptibility of hPSC-CM to chelerythrine-stimulated apoptosis varied with time after differentiation, but at most time points, hPSC-CM were more resistant than RVNC. This simple and scalable humanized high-content assay generates accurate cardiotoxicity profiles that can serve as a base for further assessment of cardioprotective strategies and drug safety.

• Authors: Maxime Mioulane, Gabor Foldes, Nadire N Ali, Michael D Schneider, Sian E Harding
• Journal: Journal of cardiovascular translational research (J Cardiovasc Transl Res) Vol. 5 Issue 5 Pg. 593-604 (Oct 2012) ISSN: 1937-5395 [Electronic] United States
• PMID: 22896035 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Benzophenanthridines
• chelerythrine
• Caspases, Effector

Topics

• Animals
• Animals, Newborn
• Apoptosis (drug effects)
• Automation, Laboratory
• Benzophenanthridines (toxicity)
• Caspases, Effector (metabolism)
• Cell Line
• Cell Membrane Permeability (drug effects)
• Cell Nucleus Shape (drug effects)
• Cell Tracking (methods)
• Dose-Response Relationship, Drug
• Enzyme Activation
• High-Throughput Screening Assays (methods)
• Humans
• Image Processing, Computer-Assisted
• Induced Pluripotent Stem Cells (drug effects, metabolism, pathology)
• Membrane Potential, Mitochondrial (drug effects)
• Microscopy, Fluorescence
• Mitochondria, Heart (drug effects, pathology)
• Myocytes, Cardiac (drug effects, metabolism, pathology)
• Necrosis
• Phenotype
• Rats
• Time Factors