Non-Invasive Image-Based Assessment of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
Year: 2014
Type: article
Abstract: ORAL ABSTRACTS S-30 Non-Invasive Image-Based Assessment of Human Induced Pluripotent Stem CeJl .. Derived Cardiomyocytes C. M. Heylman 1, R. Datta 1, Y. Kurokawa 1, D. D. Tran B. R. Conklin 2, E. Gratton 1 ~ S. C. George 1 ; University of California,. Irvine, Irvine, CA, 2 Gladstone Institutes, San Francisco, CA. ORAL ABSTRACTS S-30 S-31 ORAL ABSTRACTS Problem: Human induced pluripotent stem (iPS) cells have emerged as a viable source ofhuman cardiomyocytes for developing more accurate in vitro cardiac drug screening platforms. With improved cardiac tissue models, non-invasive, non-destructive methods that assess functional tissue metrics longitudinally in response to drugs are necessary. Objectives: The objectives of this study are to develop and val- idate non-invasive and non-destructive methods for assessing the electrophysiology and metabolic state of human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CMs). Methodology: We differentiated beating hiPS-CM from human iPS cell line WTC-11. The electrophysiological phenotype was as- sessed using the membrane voltage sensitive dye (VSD), di-4-AN- E(F)PPTEA, and the genetically encoded calcium indicator, GCaMP6. The metabolic phenotype of hiPS-CM was quantified as the ratio of glycolysis to oxidative phosphorylation using fluores- cence lifetime imaging microscopy (FL.IM) of nicotinamide adenine dinucleotide (NADH), a central metabolite. hiPS-CM were subjected to several stimuli known to affect the metabolism and/or electro- physiology of cardiomyocytes including different doses of isopro- terenol, propranolol, hypoxia, and potassium cyanide. Results: Beta blocker propranolol (!OuM) and beta adrenergic agonist isoproterenol (JOO nM) elicited a 56% decrease and a 19% increase in beat frequency, respectively. 24 hours exposure to hyp- oxia generated a shift of 17% of pixels analyzed from oxidative phosphorylation to glycolysis (p = 0.015). Potassium cyanide (4 mM) generated a significant shift of pixels analyzed from oxidative phosphorylation to glycolysis (p < 0.05). Significance: These unique electrophysiological and metabolic endpoints will facilitate the development of in vitro human cardiac tissues with the potential to identify cardiotoxic effects in drug dis- covery applications. Acknowledgments: NIH #IUH2TR000481-0l
Authors: CM Heylman, Rupsa Datta, Yosuke Kurokawa, DD Tran, BR Conklin, Éloïse Gratton, SC George
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2772189486