Abstract
Culturing atrial cells leads to a loss in their ability to be externally paced at physiological rates and to maintain their shape. We aim to develop a culture method that sustains the shape of atrial cells along with their biophysical and bioenergetic properties in response to physiological pacing. We hypothesize that adding 2,3-Butanedione 2-monoxime (BDM), which inhibits contraction during the culture period, will preserve these biophysical and bioenergetic properties. Rabbit atrial cells were maintained in culture for 24 h in a medium enriched with a myofilament contraction inhibitor, BDM. The morphology and volume of the cells, including their ability to contract in response to 1–3 Hz electrical pacing, was maintained at the same level as fresh cells. Importantly, the cells could be successfully infected with a GFP adenovirus. Action potentials, Ca2+ transients, and local Ca2+ spark parameters were similar in the cultured and in fresh cells. Finally, these cultured cells' flavoprotein autofluorescence was maintained at a constant level in response to electrical pacing, a response similar to that of fresh cells. Thus, eliminating contraction during the culture period preserves the bioelectric, biophysical and bioenergetic properties of rabbit atrial myocytes. This method therefore has the potential to further improve our understanding of energetic and biochemical regulation in the atria.
Highlights
Cardiac fibrillation is one of the leading causes of morbidity and mortality in the developed world, where atrial fibrillation (AF) is the most common sustained arrhythmia (Benjamin et al, 2017), affecting ∼2.2 million patients in the United States, and its prevalence increases with age
We aim to develop a culture method that sustains the shape of atrial cells along with their biophysical and bioenergetic properties in response to physiological pacing
The cultured cells can be infected with a GPF adenovirus after 24 h of culture. The essence of this method is the addition of a myofilament contraction inhibitor [2,3-Butanedione 2-monoxime (BDM)] to the culture medium for 24 h. This method has the potential to further improve our understanding of energetic and biochemical regulation in the atria, which can lead to novel therapies involving the adaptation of this signaling, with the goal of eliminating AF events
Summary
Cardiac fibrillation is one of the leading causes of morbidity and mortality in the developed world, where atrial fibrillation (AF) is the most common sustained arrhythmia (Benjamin et al, 2017), affecting ∼2.2 million patients in the United States, and its prevalence increases with age (one in four people over the age of 40 will develop AF; Lloyd-Jones et al, 2004). GFP gene transfection, and Förster resonance energy transfer methods have recently been developed to explore biophysical, bioelectrical and bioenergetic mechanisms (Fischer et al, 2015; Yaniv et al, 2015; Behar et al, 2016; Wüst et al, 2017). These methods are bringing new insights into AF-related molecular mechanisms, they must be applied on cells that are maintained in culture at least for 24 h. Proper measurements of physiological Ca2+ dynamics, posttranslational modification signaling, and energetic balance in cultured cells are impossible, and AF-related mechanisms cannot be explored
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