Oxygen requirements,morphology, cell coat and membrane permeability of calcium-tolerant myocytes from hearts of adult rats

Summary The morphological, functional, and biochemical properties of freshly isolated heart muscle cells were examined. A reproducible method for the separation and purification of such cells isolated from adult rat heart was developed. It yields an average of 5×10⁶ striated rectangular cells which retain normal morphology (range) 2.5 to 11×10⁶ and 4×10⁶ calcium-tolerant cells (range) 2.5 to 5.5×10⁶ per heart. After purification, 85 to 95% of the cells retain normal morphology in solutions of calcium ion activity equal to 10M, and 65 to 79% of the cells are rectangular in solutions of calcium ion activity equal to 1 mM.Under the light microscope we were able to identify functionally intact individual cells that are calcium-tolerant and contract only in response to electrical stimulation, as well as dying myocytes that beat spontaneously. The examination of such cells under the electron microscope permitted us to address the question: What is the sequence of structural changes in a dying cell? The sarcomere lengths measured both in the living state and after preparation for electron microscopy are in the physiological range. In steady states of oxygen tension, respiration of the intact cells is undiminished from 50 torr to 2 torr. The oxygen tension for half maximal respiration is 0.15 torr. Therefore, the limitation of oxygen diffusion to the mitochondria of isolated heart muscle cells must be remarkably small.This work was supported in part by NIH HL 19299 (BAW), and NIH HL 24336 and N.Y. Heart Grant in Aid (TFR.)Dr. Thomas F. Robinson is the recipient of NIH, Research Career Development Award HL 00568