Guinea pig adrenocortical cells: in vitro characterization of separated zonal cell types.

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Abstract
A quick, relatively simple and reproducible technique for obtaining populations of zona fasciculata and zona glomerulosa cells up to 80-90% pure, which can be maintained in vitro for study of adrenocortical cell function, is reported. Isolated guinea pig adrenocortical cells were separated on a 1-28% bovine serum albumin/Ca2+, Mg2+-free buffer gradient (wt/vol at 4% increments) using equilibrium density centrifugation (570 g, 30 min). Over 60% of the 8 .times. 105 viable cells/adrenal obtained in the total isolate were recovered after separation. Of the zona glomerulosa cells, 80% were found in the lower 3 bands of the gradient. Of the zona fasciculata cells, 78% were found in the top 3 bands. Of the cells in the first 2 bands, 78-91% were zona fasciculata cells, while, of the cells in the bottom two bands, 92-95% were zona glomerulosa cells. The cells retained the morphological characteristics of cells in situ and could be maintained in vitro for periods up to 11 days. They produced a wide variety of steroids, cortisol, corticosterone, aldosterone, 11-.beta.-hydroxyandrostenedione, deoxycortisol, deoxycorticosterone, cortisone, 18-hydroxycorticosterone, and a product tentatively identified as dehydroepiandrosterone, and they responded to ACTH in a dose-responsive manner with enhanced levels of steroid output. Zona glomerulosa-enriched populations differed from zona fasciculata-enriched populations in their abundant production of aldosterone and in the pattern of steroid production. None of the cultures responded to angiotensin II (100 pg/ml) with increased steroid production.