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At an unknown point in mammalian development, cardiac precursor cells become committed to the cardiocyte phenotype. Certain of these are believed to specialize further into pacemaker cardiocytes. By culturing explanted embryonic ventricles into in vivo organ culture (Tucker DC, Snider C, Woods WT Jr: Pediatr Res 23:637-642, 1988), we observed pacemaker cells arising apparently from cardiocytes. We hypothesized that this event can be triggered by intercellular attachments, innervation, vascularization, or other factors. The present study was designed to test the hypothesis that primitive ventricular cells in the tubular heart can organize into an anatomically and electrophysiologically distinct pacemaker structure in the absence of innervation or vascularization from extrinsic sources. Developing ventricles of tubular hearts from 10-d-old rat embryos (n = 22) were excised and incubated in culture dishes. Within each explant, a group of cells organized into a uniform cluster (diameter, 150 +/- 50 microns) after 8 +/- 2 d in culture. These cells resembled immature conduction system cells and had electrophysiologic features similar to those of mature pacemaker cells. Single-cell action potentials and impulse conduction patterns proved that the pacemaker cluster was the dominant pacemaker for the entire explant. These results confirm that, in the absence of extrinsic nerves and blood vessels, culture of the tubular ventricle elicits formation of an organized pacemaker, resembling the mature cardiac pacemaker and suggesting that contact with the culture dish surface or some other factor triggers conversion of cardiocytes to the pacemaker phenotype.

Organization and Fine Structure of a Pacemaker Derived from Fetal Rat Myocardium