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Cardiovascular Research 2003 58(2):423-434; doi:10.1016/S0008-6363(03)00253-0
© 2003 by European Society of Cardiology
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Copyright © 2003, European Society of Cardiology

The human adult cardiomyocyte phenotype

S.D. Birda, P.A. Doevendansb,c,*, M.A. van Rooijena, A. Brutel de la Riviered, R.J. Hassinka,d, R. Passiera,b and C.L. Mummerya,b

aHubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht, The Netherlands
bInteruniversity Cardiology Institute of the Netherlands, Utrecht, The Netherlands
cDepartment of Cardiology, Heart Lung Center, Utrecht, The Netherlands
dCardiothoracic Surgery, Heart Lung Center, Utrecht, The Netherlands

p.doevendans{at}hli.azu.nl

* Corresponding author. Present address: Department of Cardiology, HLCU, UMC, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands. Tel.: +31-30-250-9111; fax: +31-30-254-2155.

Aim: Determination of the phenotype of adult human atrial and ventricular myocytes based on gene expression and morphology. Methods: Atrial and ventricular cardiomyocytes were obtained from patients undergoing cardiac surgery using a modified isolation procedure. Myocytes were isolated and cultured with or without serum. The relative cell attachment promoting efficiency of several reagents was evaluated and compared. Morphological changes during long-term culture were assessed with phase contrast microscopy, morphometric analysis and immunocytochemistry or RT-PCR of sarcomeric markers including {alpha}-actinin, myosin light chain-2 (MLC-2) and the adhesion molecule, cadherin. Results: The isolation method produced viable rod-shaped atrial (16.6±6.0%, mean±S.E.; n = 5) and ventricular cells (22.4±8.0%, mean±S.E.; n = 5) in addition to significant numbers of apoptotic and necrotic cells. Cell dedifferentiation was characterized by the loss of sarcomeric structure, condensation and extrusion of sarcomeric proteins. Cells cultured with low serum recovered and assumed a flattened, spread form with two distinct morphologies apparent. Type I cells were large, had extensive sarcolemmal spreading, with stress fibers and nascent myofibrils, whilst type II cells appeared smaller, with more mature myofibril organisation and focal adhesions. Conclusion: Characterization of the redifferentiation capabilities of cultured adult cardiac myocytes in culture, provides an important system for comparing cardiomyocytes differentiating from human stem cells and provides the basis for an in vitro transplantation model to study interaction and communication between primary adult and stem cell-derived cardiomyocytes.

KEYWORDS Adult human cardiomyocyte; Cell isolation; Cell attachment; Laminin; Myofibrillogenesis; Sarcomere; Cadherin


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