The contractor shall supply NICHD investigators with freshly prepared or cultured rat pituitary cell preparations, and freshly-prepared rat adrenal preparations. The contractor shall prepare viable hormone-responsive cells from rat pituitary and adrenal glands in the following conditions and quantities: (a) Pituitary glands: anterior pituitary glands from 50 adult female rats shall be obtained three times weekly, finely minced, and dispersed into isolated cells by enzymatic digestion with trypsin and DNAse. The acutely dispersed cells will be cultured for 2 days in culture plates or plastic dishes under conditions specified by the Project Officer, usually at one million per 35 mm will, prior to delivery to the laboratory. (b) Adrenal glands: Adrenal gland from 50 rats will be removed thrice weekly and separated into capsular (glomerulosa) and inner (fasciculata-reticularis) zones prior to enzyme treatment. The capsules will be minced and digested with collagenase to prepare viable glomerulosa cell dispersions with the capacity to respond to peptide hormones by aldosterone production in vitro.
