The Isolated Bovine Adrenomedullary Chromaffin Cell: A Model of Neuronal Excitation-Secretion

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Abstract
A method is described for the isolation of functional chromaffin cells from bovine adrenal medulla. The procedure involves dispersion of minced medullary tissue with a mixture of crude collagenase and hyaluronidase in calcium-free medium, combined with constant movement of the mince over nylon cloth. Dispersed cells were separated from larger tissue fragments by filtration through nylon cloth and glass filters of successively decreasing mesh size. Cells were collected and simultaneously removed from cell fragments and dispersion medium by centrifugation through a concentrated solution of bovine serum albumin. The isolated cells have been identified as chromaffin cells by a histochemical fluorescence technique which converts intracellular catecholamines to fluorescent condensation products with gaseous formaldehyde. This method has been combined with subsequent Giemsa staining of the smears to demonstrate that less than 1% of contaminating cells are present. Isolated cells were viable as indicated by exclusion of the dye trypan blue and by secretion of catecholamines in response to stimulation with acetylcholine in the presence of neostigmine. It is suggested that the isolated chromaffin cell might provide a simple model system for studying the processes which couple excitation and secretion in neurons.