Effect of a simple versus a complex matrix on the polarity of cardiomyocytes in culture.

Abstract

Objective: The objective of this study was to observe the effects of cell culture on cellular polarity in cardiomyocytes as influenced by cytoskeletal proteins.

Methods: Cardiomyocytes from adult and neonatal rats were isolated and grown on 2 different extracellular matrices--laminin and a complex, fibroblast-derived extracellular matrix, cardiogel. The location of a number of proteins was visualized by means of fluorescence deconvolution microscopy, using specific fluorescent probes for alpha-adrenergic receptors, beta-adrenergic receptors, the sarcolemmal L-type calcium channel, and the sodium + potassium adenosine triphosphatase pump protein. Intracellular migration of these proteins during the first 4 days of culture was followed and microscopic stacked images were reconstructed. A fluorescein isothiocyanate-labeled probe for actin was used to ensure that cardiomyocytes were being examined, based on protein patterns.

Results: We examined 2 types of myocyte: freshly isolated neonates and cultured adult cardiomyocytes that undergo dedifferentiation. Initial, perinuclear clumping (endoplasmic reticulum/Golgi-associated) of the probes with an ensuing spread to the cytoplasm and periphery, accompanied by a better organization and more rapid response to biochemical stimuli, was seen on the complex matrix.

Conclusions: A complex matrix overcomes cell polarity at a faster rate than myocytes cultured on a simple matrix, although both culture matrices were able to support cell growth and differentiation, and single-layer cultures are a good method by which structural and biochemical data can be obtained. The use of a native, complex matrix is preferable to employing a simple, single protein, although temporal aspects of cell growth must be considered regarding the particular aspect of the cell structure development/biochemical pathways that the researcher intends focusing on.