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Chemotherapy-induced impairment of autophagy is implicated in cardiac
toxicity induced by anti-cancer drugs. Imperfect translation from rodent
models and lack of in vitro models of toxicity has limited investigation
of autophagic flux dysregulation, preventing design of novel
cardioprotective strategies based on autophagy control. Development of an
adult heart tissue culture technique from a translational model will
improve investigation of cardiac toxicity. We aimed to optimize a canine
cardiac slice culture system for exploration of cancer therapy impact on
intact cardiac tissue, creating a translatable model that maintains
autophagy in culture and is amenable to autophagy modulation. Canine
cardiac tissue slices (350 μm) were generated from left ventricular free
wall collected from euthanized client-owned dogs (n=7) free of
cardiovascular disease at the Foster Hospital for Small Animals at Tufts
University. Cell viability and apoptosis were quantified with MTT assay
and TUNEL staining. Cardiac slices were challenged with doxorubicin and an
autophagy activator (rapamycin) or inhibitor (chloroquine). Autophagic
flux components (LC3, p62) were quantified by western blot. Cardiac slices
retained high cell viability for >7 days in culture and basal
levels of autophagic markers remained unchanged. Doxorubicin treatment
resulted in perturbation of the autophagic flux and cell death, while
rapamycin co-treatment restored normal autophagic flux and maintained cell
survival. We developed an adult canine cardiac slice culture system
appropriate for studying the effects of autophagic flux that may be
applicable to drug toxicity evaluations.
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