ウシ副腎皮質細胞の初代培養法

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The isolated bovine adrenocortical cells are prepared aseptically by the use of collagenase and deoxyribonuclease. The isolated cells are suspended in Ham F-10 medium containing 5% fetal calf serum, 10% newborn calf serum, 2.5% horse serum and antibiotics. The seeded cells are cultured at 37°C in a humidified atmosphere of 5% CO<SUB>2</SUB> in air. Steroidogenic activity for ACTH reached the maximum in the 2 to 3-day primary cultured cells; the maximum response to ACTH in these cells is more intense than that in freshly isolated bovine adrenocortical cells. The primary cultured cells have prostaglandin, muscarinic, ATP and β-adrenergic receptors that are linked to steroidogenesis in addition to ACTH and aldosterone receptors. Thus primary cultured bovine adrenocortical cells are a useful tool to study these receptors and the intracellular events that are associated with the receptors. We also demonstrated that the fura 2 loaded primary cultured monolayer cells on glass cover slips provide us much more information than suspended cells in the study of intracellular Ca<SUP>2+</SUP> mobilization in adrenocortical cells.
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