Abstract
In this work we demonstrate a label-free optical imaging technique to assess metabolic status and oxidative stress in human induced pluripotent stem cell-derived cardiomyocytes by two-photon fluorescence lifetime imaging of endogenous fluorophores. Our results show the sensitivity of this method to detect shifts in metabolism and oxidative stress in the cardiomyocytes upon pathological stimuli of hypoxia and cardiotoxic drugs. This non-invasive imaging technique could prove beneficial for drug development and screening, especially for in vitro cardiac models created from stem cell-derived cardiomyocytes and to study the pathogenesis of cardiac diseases and therapy.
Highlights
Many drugs showing promise in preclinical trials fail during clinical development due to the emergence of cardiac side effects [1,2,3]
We demonstrate how fluorescence lifetime imaging microscopy (FLIM) imaging can be used to characterize the metabolic state and oxidative stress in human induced pluripotent stem cell-derived cardiomyocyte (hiPS-CM) using phasor analysis of endogenous biomarkers
Our results demonstrate the potential of this technique to detect drug-induced stress and pathological conditions in human induced pluripotent stem (hiPS)-CMs
Summary
Many drugs showing promise in preclinical trials fail during clinical development due to the emergence of cardiac side effects [1,2,3]. The development of in vitro platforms that accurately mimic the biology of human cardiac cells provide a plausible model for high-throughput drug screening to detect potential cardiotoxicity before wide spread human use. The emergence of human induced pluripotent stem cell technology has expanded the possibilities for sourcing human cardiomyocytes [4,5]. There is a need for non-destructive techniques to visualize the biology and drug response of such in vitro human induced pluripotent stem cell-. Derived cardiomyocyte (hiPS-CM) models for drug screening. Indices of metabolism and oxidative stress are relevant in cardiac pathologies and response to drugs [6,7]. We employ two-photon fluorescence lifetime imaging microscopy (FLIM) to assess the metabolic state and oxidative stress in hiPS-CMs
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