An Improved Technique for the Preparation of Isolated Rat Adrenal Cells: A Sensitive, Accurate and Specific Method for the Assay of ACTH

Hypothalamic peptides influencing secretion of ACTH by isolated adenohypophysial cells

“…The cells were incubated at 37'C for 60 min in a Dubnoff shaker, under an atmosphere of 95% Os, 5% COz. The media were separated by centrifugation (800 X g for 10 min), acidified to pH 3.5 with HCl and assayed for ACTH by the isolated adrenal cortex cell technique [3]. Fraction CRF,(G-15) is our interim house standard; one corticotrophin unit is 236 the activity exhibited by 5 pg of this material.…”

Effect of Temperature during Preparation of Rat Adrenal Quarters and Isolated Cell Suspension on Their ACTH Responsiveness

“…Immediately Isolated adrenal cell suspension was prepared, using a modification of the method of Sayers et al (1971). Adrenal gland was cut into 4 pieces at room temperature.…”

“…for ACTH in this experiment was a solution of 0.5% bovine serum albumin in 0.9% NaCl, pH being adjusted to 3.5 with 0.1N HCl as described by Sayers et al(1971). The tubes were flushed with a gas …”

Acth and MSH Assays and Their Clinical Application

“…Quartered rat adrenal glands (Saffron & Schally, 1955) or suspensions of separated adrenocortical cells (Kloppenberg et al, 1968;Swallow & Sayers, 1969;Sayers et al, 1971), maintained in vitro, are sensitive to ACTH; the fluorogenic corticosteroid production is used as the index of response. These assay systems are technically easier to perform than the in vivo systems, and the isolated cell method is much more sensitive, a minimum detectability of 0.1-1 pU (1-10 pg) ACTH being achieved (Sayers el al., 1971).…”