A Trypsin Technic for the Preparation of Isolated Rat Anterior Pituitary Cells

Oxytocin at physiological concentrations evokes adrenocorticotropin (ACTH) release from corticotrophs by increasing intracellular free calcium mobilized mainly from intracellular stores. Oxytocin displays synergistic or additive effects on ACTH-releasing factor or arginine vasopressin-induced ACTH secretion, respectively.

“…Anterior pituitary cells were isolated as described previously (27)(28)(29). Briefly, the excised adenohypophyses were placed in medium A (Dulbecco's MEM, buffered with 22 mM HEPES at pH 7.4, containing 0.25% BSA and antibiotic-antimycotic substances: Penicillin G, 100 U/ml; Streptomycin, 100 ~g/ml; Amphotericin B, 0.25 ~g/ml) and minced to approximately 0.5 mm diameter segments.…”

An autoradiographic study of the intracellular transport of interstitial cell stimulating hormone in dissociated pituitary glands of normal and castrated rats

“…The first was that of Hopkins and Farquhar (1973); the second was the technique Portanova et al (1970) had used. The method used to dissociate the tissue in this study employs some procedures from each of the studies men tioned above.…”

“…As far as junctions are concerned, the total area of cell surfaces devoted to them is limited in anterior pituitary cells compared to the situation encoun tered in the liver, exocrine pancreas, thyroid gland, intes tine, and other lumen-lining epithelia (Farquhar et al, 1975). A number of successful attempts have been made to pro duce viable single cell suspensions from a variety of tis sues, e.g., liver (Howard et al, 1967;Berry and Friend, 1969), adrenal gland Halkerston et al, 1968;Swallow and Sayers, 1969), corpus luteum (Gospodarowicz and Gospodarowicz, 1972), thyroid gland (Maayan and Ingbar, 1968) and exocrine pancreas (Amsterdam and Jamieson, 1972) including the anterior pituitary (Portanova et al, 1970;Hymer and Evans, 1970;Bala et al, 1970;Sayers et al, 1971;Vale et al, 1972;Ishikawa, 1969;Leavitt et al, 1973;and Hymer et al, 1973). Most of these procedures have involved the use of collagenase, hyaluronidase, trypsin, pronase, EDTA, and calcium-free media, alone or in various com binations.…”

“…However there were still some silver grains over the endoplasmic reticulum The dissociation technique used in this study for the preparation of rat anterior pituitary cells is adapted from techniques previously reported by Hopkins and Farquhar (1973) and Portanova et al (1970). Both of these techniques pro vided large numbers of dispersed cells.…”

“…One procedure was that used by Hopkins and Farquhar (1973) which employs the subsequent incubation of tissue blocks in trypsin, DNAase, soya bean trypsin inhibitor, EDTA and neuraminidase followed by gentle shearing to disperse the tissue blocks. The other method, used by Portanova et al (1970), utilizes mechanical agitation in trypsin with subsequent washing in lima bean trypsin inhibitor to obtain suspensions of isolated anterior pituitary cells.…”