Primary tissue culture of normal adult rabbit adrenocortical cells fromzonae fasciculata andreticularis

Abstract

SUMMARY: Parenchymal cells isolated from decapsulated adrenal cortices of adult New Zealand white female rabbits by means of enzymatic {trypsin plus collagenase and hyaluronidase) predigestion at 37 ° C followed by mechanical dissociation within a calcium (Ca~+)-free buffer were set up in primary monolayer cultures onto very thin, porous polyethylene discs floating on the top of a growth medium contained inside the wells of a plastic cluster plate. In this setting, adrenal cortical cells, being at the interface between air and growth medium, benefit from both the respiratory and metabolic standpoint. Therefore, they fully respond to anabolic and growthpromoting stimuli which may increase cellular oxygen consumption significantly. Hence, this in vitro model is well suited for studying the fairly complex regulatory mechanisms of mammalian adult adrenal cortical cell proliferation and differentiation, as it allows one to take full advantage of an artificially controlled environment and of completely synthetic growth media.

Extracellular calcium modulates prereplicative cyclic AMP surges in EGF‐stimulated primary neonatal rat Hepatocytes

“…The HiWo,Ba,,,, medium was either Ca2+-free or contained 10 mgil CaCl, and contained 7.5 mgil ornithine instead of arginine (Menapace and Armato, 1987). Just before use, 50 IU/ml streptomycin (Squibb and Sons S.p.A., Rome), 5.35 pg/ml imipenem monohydrate (Sigma Tau S.p.A., Rome), 5.35 pg/ml sodium cilastatin (Sigma Tau), and 250 ngiml fungizone (Mascia-Brunelli S.p.A., Milan) were added to the medium.…”