Structural and elemental characterization of heart cells grown in a collagen matrix

Abstract

A novel preparation of spontaneously contracting heart cells embedded in a collagen strand provides an ideal experimental system for correlative structure-function experiments that utilize the techniques of electron microscopy, quantitative electron probe x-ray microanalysis (EPXMA) and imaging. Heart cells grown within the strand for 1 day possess the subcellular content and distribution of physiologically relevant elements—Na, Mg, P, 5, Cl, K, and Ca—found in intact heart cell preparations. The presence of junctional specializations between, and organized myofibrils within, the majority of cells after 1 day in culture also establishes that the collagen matrix promotes vigorous cell development as well as maintains physiological integrity. EPXMA, combined with ultrastructural analyses, provides elemental content data on a cell-by-cell basis. In studies presented here, viable cells, comprising over 80% of the strand cell population, could be distinguished easily from those which had been functionally compromised, not only by aberrant structure but also by altered subcellular compartmentation of Na, K, Cl, and Ca. Within individual viable cells, compartmental differences in element content were notable especially between mitochondria and cytoplasm. However, nuclear euchromatin, but not heterochromatin, appeared approximately identical to cytoplasm in elemental and water content. In such cells, the cytoplasmic K:Na ratio was maintained at a high level (∼15:1). The results with respect to K, Na, and other elements demonstrated the integrity of membrane transport mechanisms regulating the movement and distribution of ions and the maintenance of ionic homeostasis in cells of the strand preparation.