Primary mouse pituitary cell culture

Whole pituitaries were dissected from wild-type C57BL/6 female mice at 8 to 9 weeks of age. Mice were cohoused but not assessed for estrous cycle stage at harvest. Mice were handled in accordance with University of California, San Diego, Institutional Animal Care and Use Committee regulations under an approved protocol. Whole pituitaries were isolated in ice-cold PBS and then dispersed by incubation with collagenase (Sigma Aldrich, St. Louis, MO) and 0.25% trypsin (Sigma-Aldrich) as previously described (5). After plating on poly-l-lysine (Sigma-Aldrich)‒coated four-well Nunc Lab-Tek II chamber slides (Thermo Fisher Scientific, Waltham, MA), the cells were allowed to recover for at least 48 hours at 37°C and 5% CO₂ in high-glucose HEPES-buffered DMEM with 10% FBS before experimentation. No exogenous steroids were provided in the culture medium.