Journal
EXPERIMENTAL CELL RESEARCH
Volume 317, Issue 2, Pages 188-194Publisher
ELSEVIER INC
DOI: 10.1016/j.yexcr.2010.08.017
Keywords
Cardiomyocyte renewal; Ploidy; Pericentriolar material 1; Cardiac Troponin; lododeoxyuridine
Categories
Funding
- Swedish Research Council
- Swedish Cancer Society
- Karolinska Institute
- Tobias Stiftelsen
- Knut och Alice Wallenbergs Stiftelse
- Deutsche Forschungsgemeinschaft
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Assays to quantify myocardial renewal rely on the accurate identification of cardiomyocyte nuclei. We previously C-14 birth dated human cardiomyocytes based on the nuclear localization of cTroponins T and I. A recent report by Kajstura et al. suggested that cTroponin I is only localized to the nucleus in a senescent subpopulation of cardiomyocytes, implying that C-14 birth dating of cTroponin T and 1 positive cell populations underestimates cardiomyocyte renewal in humans. We show here that the isolation of cell nuclei from the heart by flow cytometry with antibodies against cardiac Troponins T and I, as well as pericentriolar material 1 (PCM-1), allows for isolation of close to all cardiomyocyte nuclei, based on ploidy and marker expression. We also present a reassessment of cardiomyocyte ploidy, which has important implications for the analysis of cell turnover, and iododeoxyuridine (IdU) incorporation data. These data provide the foundation for reliable analysis of cardiomyocyte turnover in humans. (C) 2010 Elsevier Inc. All rights reserved.
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