Isolation of Cardiomyocytes from Fixed Hearts for Immunocytochemistry and Ploidy Analysis.

ABSTRACT: The adult mammalian heart is composed of various cell types including cardiomyocytes, endothelial cells and fibroblasts. Since it is difficult to reliably identify nuclei of cardiomyocytes on histological sections, many groups rely on isolating viable cardiomyocytes prior to fixation to perform immunostaining. However, these live cardiomyocyte isolation techniques require optimization to maximize the yield, viability and quality of the samples, with inherent fluctuations from sample to sample despite maximum optimization. Here, we report a reproducible protocol, involving fixation prior to enzymatic digestion of the heart, which leads to maximum yield while preserving the in vivo morphology of individual cardiomyocytes. We further developed an automated analysis platform to determine the n

SUBMITTER: Yucel D 

PROVIDER: S-EPMC7993010 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature