Isolation of cardiac myocytes from the adult newt, Notophthalmus viridescens. an electron microscopic and quantitative light microscopic analysis.
This report describes a technique which permits a high yield of viable adult cardiac myocytes from the adult newt using enzymatic separation techniques at low temperature and high enzyme concentrations. Observations by light microscopy showed the isolated myocytes to have a distinctively slender morphology which consisted of a variable number of arm-like appendages radiating from the center of cells which were predominantly mononucleated. Atrial myocytes were typically observed to have two to three arm-like appendages while ventricular myocytes typically had three to six appendages. The majority of myocytes displayed normal fine structure when examined by transmission electron microscopy. Computerized image analysis revealed that atrial cells were significantly greater in cell length (192.9 +/- 53.4 microns) and in nuclear length (25 +/- 5.3 microns) and perimeter (59.2 +/- 10.7 microns) than were ventricular cells (162.8 +/- 39 microns, 23.6 +/- 5.1 microns and 57.4 +/- 11.1 microns, respectively), while cell widths and areas were greater in ventricular cells (16.5 +/- 4.7 microns and 1839.8 +/- 585.0 microns, respectively) than in atrial cells (13.2 +/- 3.1 microns and 1520.3 +/- 527.6 microns, respectively). Comparison of these data with previous descriptions of isolated amphibian and mammalian cardiac myocytes emphasizes species-related differences.
D000818:Animals; D002469:Cell Separation / Q000379:methods; D008854:Microscopy, Electron; D009206:Myocardium / Q000166:cytology* / Q000648:ultrastructure; D012455:Salamandridae