Isolation and Use of Primary Adrenocortical Cells from Guinea Pigs, Dogs and Monkeys forIn Vitro Toxicity Studies

Abstract

A method was developed to obtain enriched populations of zona fasci-culata cells from the adrenal glands of guinea pigs, dogs, and monkeys. Adrenocortical cells (ADC) in primary culture were shown to maintain viability and cellular morphology, with an estimated ≥80% of cultures being zona fasciculata cells. Three known adrenal toxicants, PD 132301–2, 1-(o-chlorophenyl)-1-(p-chloro-phenyl)-2, 2-dichloroethane (o, p′-DDD), and aminoglutethimide (AG) were tested in this in vitro system. Neutral red (NR) uptake was used as a marker of cell viability and cortisol production was measured to assess ADC function. NR uptake following 24 h of treatment with PD 132301–2 (10 μM) was 32, 31, and 53% of control in guinea pig, dog, and monkey cultures, respectively. Similarly, o, p'-DDD (100 μM) decreased NR uptake to 32, 40, and 69% of control. AG (300 μM) decreased NR uptake by 50% only in dog ADC. Cortisol production was evident in cells from all three species with rates being highest in monkey, followed by the dog and guinea pig. Cortisol production was decreased following treatment with all three toxicants. Decreases paralleled loss of viability in PD 132301–2-treated cultures from all three species and in o, p'-DDD-treated cultures from guinea pig and dog. In contrast, decreased cortisol production preceded any change in viability in o, p'-DDD-treated cultures from monkey, and in AG-treated cultures from all species. Cytotoxic responses to adrenal toxicants of varied structure and mechanisms of action suggests that the ADC cultures from these three species may be useful in toxicologic screening or for investigating mechanisms of adrenocortical toxicity. Key Words: Adrenocortical cells—-Cytotoxicity—Cortisol—o, p'DDD—Aminoglutethimide—PD 132301–2.