Isolation of Cardiomyocytes and Cardiofibroblasts for Ex Vivo Analysis.
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.
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia.
George Williams
GW
Mbogo
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia.
Christina
C
Nedeva
Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, VIC, 3086, Australia. H.puthalakath@latrobe.edu.au.
Hamsa
H
Puthalakath
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis; D002118:Calcium / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D006333:Heart Failure / Q000503:physiopathology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism
D002118:Calcium
Apoptosis; Cardiofibroblasts; Cardiomyocytes; Ex vivo models; Heart failure