Abstract
Heart failure (HF) is a common clinical endpoint to several underlying causes including aging, hypertension, stress, and cardiomyopathy. It is characterized by a significant decline in the cardiac output. Cardiomyocytes are terminally differentiated cells and therefore, apoptotic death due to beta adrenergic (β-AR) signaling contributes to high attrition rate of these cells. Past treatments of HF offer some survival benefit to patients (e.g., the beta blockers), but at the expense of blocking the compensatory beta-adrenergic signaling in surviving cells. One prerequisite for developing new therapeutics is to be able to grow cardiomyocytes ex vivo, and test their apoptotic response to drugs. Here we describe methods for isolation and culturing of neonatal and adult calcium tolerant cardiomyocytes. Similarly, cardiofibroblasts can also be isolated using the same protocol and subsequently, immortalized with SV40 T-Antigen for ex vivo studies.
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Acknowledgements
This work was supported by the National Health and Medical Research Council Grant No. 1085281 to HP. GWM and CN are supported by La Trobe University Post graduate Research scholarships.
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Mbogo, G.W., Nedeva, C., Puthalakath, H. (2016). Isolation of Cardiomyocytes and Cardiofibroblasts for Ex Vivo Analysis. In: Puthalakath, H., Hawkins, C. (eds) Programmed Cell Death. Methods in Molecular Biology, vol 1419. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3581-9_10
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DOI: https://doi.org/10.1007/978-1-4939-3581-9_10
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