ISOLATION AND USE OF PRIMARY ADRENOCORTICAL-CELLS FROM GUINEA-PIGS, DOGS AND MONKEYS FOR IN-VITRO TOXICITY STUDIES

A method was developed to obtain enriched populations of zona fascicul ata cells from the adrenal glands of guinea pigs, dogs, and monkeys. A drenocor tical cells (ADC) in primary culture were shown to maintain v iability and cellular morphology, with an estimated greater than or eq ual to 80% of cultures being zona fasciculata cells. Three known adren al toxicants, PD 132301-2, hlorophenyl)-1-(p-chlorophenyl)-2,2-dichlor oethane (o,p'-DDD), and aminoglutethimide (AG) were tested in this in vitro system. Neutral red (NR) uptake was used as a marker of cell via bility and cortisol production was measured to assess ADC function. NR up- take following 24 h of treatment with PD 132301-2 (10 mu M) was 3 2, 31, and 53% of control in guinea pig, dog, and monkey cultures, res pectively. Similarly, o,p-DDD (100 mu M) decreased NR uptake to 32, 40 , and 69% of control. AG (300 mu M) decreased NR uptake by 50% only in dog ADC. Cortisol production was evident in cells from all three spec ies with rates being highest in monkey, followed by the dog and guinea pig. Cortisol production was decreased following treatment with all t hree toxicants. Decreases paralleled loss of viability in PD 132301-2- treated cultures from all three species and in o,p'-DDD-treated cultur es from guinea pig and dog. In contrast, decreased cortisol production preceded any change in viability in o,p'-DDD-treated cultures from mo nkey, and in AG-treated cultures from all species. Cytotoxic responses to adrenal toxicants of varied structure and mechanisms of action sug gests that the ADC cultures from these three species may be useful in toxicologic screening or for investigating mechanisms of adrenocortica l toxicity.