Characterization of Two Chromaffin Cell Populations Isolated from Bovine Adrenal Medulla

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Abstract
Cultures of chromaffin cells isolated from the bovine adrenal medulla have been extremely useful for investigating secretory mechanisms, but such cultures used up to the present time represent mixed populations of adrenergic and noradrenergic cells. This report describes how, with slight modifications to standard procedures, two separate chromaffin cell populations may be separated from bovine adrenal medullae. These two cell fractions have been characterized by biochemical, immunocytochemical, and morphological techniques as enriched populations of adrenergic or noradrenergic cells, respectively. The adrenergic cell-enriched fraction consists of greater than 90% adrenergic cells, whereas the noradrenergic cell-enriched fraction contains greater than 60% noradrenergic cells. We also demonstrate that these cells may be cultured with their secretory machinery intact: analysis of secreted catecholamines from nicotine- or high K+ concentration-stimulated cells cultured from each fraction confirms that adrenaline is the major catecholamine secreted by one fraction, whereas noradrenaline is mainly secreted by the other.