Isolation and fractionation in percoll gradient of guinea pig adrenal cortex cells and their functional characteristics

1989 
Suspension of isolated adrenal cells from the guinea pig adrenals was prepared using two modifications of collagenase dispersion. The cells separated by percoll density gradient gave 4 bands. The fraction of adrenocorticocytes has buoyant density of 1.03-1.05 g/ml. The lower bands comprise blood cells with density of 1.07-1.10 g/ml. The aldosterone content in the adrenocorticocyte fraction is 1200-1300 pg/10(5) cells. Intensity of labelling of cellular proteins increases only in the adrenocorticocyte fraction in response to a rise in K+ concentration in incubation medium. The Scatchard plot method was used to characterize binding of 125I-ACTH to dispersed adrenocortical cells. Binding association constant (Ka) and binding capacity (Bmax) are 1.3 X 10(8) M-1 and 2.0 X 10(-9) mol/l, respectively. A relation between functional activity of adrenocortical cells, their ultrastructural features and density is discussed.
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