Studies of isolated adult rat heart cells: the surface morphology and the influence of extracellular calcium ion concentration on cellular viability.
Rat cardiac muscle was dissociated into single cells by a coronary perfusion technique with collagenase and hyaluronidase in a Ca-free medium. Retention of the cylindrical shape of isolated muscle cells could be achieved by regulation of [Ca2+]0 and temperature. Cells kept at 4 degrees C, and 0-01 mM CaCl2 remained cylindrical for more than a week and contracted spontaneously upon warming at 37 degrees C. At [Ca2+]0 between 0-1-2 mM and 37 degrees C, cells underwent contracture and rounded up. Scanning (SEM) and transmission electron microscopy were used to analyze the structure of cylindrical and rounded muscle cells. The extracellular aspect of the sarcolemma at lateral cell surfaces and intercalated disc regions were clearly revealed for SEM analysis. Both the distribution and number of T-tubule openings on the surfaces can be estimated and a three-dimensional description of the intercalated disc obtained. This study reveals that isolated adult heart cells are extremely sensitive to [Ca2+]0, but with careful control of this cation, this preparation should be helpful in the analysis of both sarcolemmal structure and the pathological changes which accompany myocardial injury.