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Cell Cycle. 2014;13(6):910-8. doi: 10.4161/cc.27768. Epub 2014 Jan 14.

Characterization of contracting cardiomyocyte colonies in the primary culture of neonatal rat myocardial cells: a model of in vitro cardiomyogenesis.

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Center of Cytoanalysis; Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences; St. Petersburg, Russia; Institute of Experimental Medicine; Federal Almazov Medical Research Centre; St. Petersburg, Russia.
Institute of Experimental Medicine; Federal Almazov Medical Research Centre; St. Petersburg, Russia.


The unmet clinical need for myocardial repair after irreversible ischemic injury requires a better understanding of the biological properties of cardiac stem cells (CSCs). Using a primary culture of neonatal rat myocardial cells, we describe the formation and maturation of contracting cardiomyocyte colonies stemming from c-kit(+), Sca(+), or Isl1(+) CSCs, which occurs in parallel to the hypertrophy of the major cardiac myocyte population. The contracting cardiomyocyte colonies (~1-2 colonies per 1 × 10(5) of myocardial cells) were identified starting from eighth day of culturing. At first, spontaneous weak, asynchronous, and arrhythmic contractions of the colonies at a rate of 2-3 beats/min were registered. Over time, the contractions of the colonies became more synchronous and frequent, with a contraction rate of 58-60 beats/min by the 30th day of culturing. The colonies were characterized by the CSCs subtype-specific pattern of growth and structure. The cells of the colonies were capable of spontaneous cardiomyogenic differentiation, demonstrating expression of both sarcomeric α-actinin and α-sarcomeric actin as well as the maturation of contractile machinery and typical Ca(2+) responses to caffeine (5 mМ) and K(+) (120 mМ). Electromechanical coupling, characterized by cardiac muscle-specific Ca(2+)-induced Ca(2+) release, was evident at 3 weeks of culturing. Thus, the co-cultivation of CSCs with mature cardiac cells resulted in the formation of contracting cardiomyocyte colonies, resembling the characteristics of in vivo cardiomyogenesis. The proposed model can be used for the investigation of fundamental mechanisms underlying cardiomyogenic differentiation of CSCs as well as for drug testing and/or other applications.


Ca2+-induced Ca2+ release; cardiac myocytes; contracting cardiomyocyte colonies; differentiation; electromechanical coupling; primary culture of neonatal myocardial cells; proliferation; resident cardiac stem cells

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