Levay-Young B K, Bandyopadhyay G K, Nandi S
Cancer Research Laboratory, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1987 Dec;84(23):8448-52. doi: 10.1073/pnas.84.23.8448.
A two-step culture system has been developed to analyze the role of hormones in casein accumulation by mammary epithelial cells obtained from adrenalectomized and ovariectomized adult virgin mice. In the first step cells are grown inside collagen gel in medium containing insulin, epidermal growth factor (EGF), and linoleic acid for 9 days; these conditions stimulate very little casein accumulation. Following this growth phase the gels are released to float in medium containing insulin, prolactin, and linoleic acid. During this second phase the mammary cells will accumulate large amounts of casein, but only in the simultaneous presence of insulin, prolactin, and linoleic acid; in the absence of linoleic acid casein accumulation is greatly reduced. The casein accumulation is not dependent on the presence of the glucocorticoid cortisol and will occur in spite of the presence of the antiglucocorticoid agent RU 38 486. To determine if the response to cortisol observed in organ culture by other investigators might be mediated by stromal cells, epithelial cells were grown in collagen gel under fatty acid-free conditions and then cocultured with explants of mammary fat pads from adult virgin mice with or without mammary parenchyma. The cocultures were performed in fatty acid-free medium containing insulin and prolactin with or without cortisol. In the majority of experiments the mammary epithelial cells in the collagen gel accumulate more casein in the presence of cortisol than in its absence, irrespective of the presence of mammary parenchyma in the explant. Thus, mammary epithelial cells are directly dependent on insulin and prolactin for casein accumulation and indirectly dependent on cortisol by means of its effect on the stromal cells. This cortisol effect may be to cause release into the medium of linoleic acid or a metabolic product of linoleic acid from the stromal cells.
已开发出一种两步培养系统，用于分析激素在取自肾上腺切除和卵巢切除的成年处女小鼠的乳腺上皮细胞酪蛋白积累中的作用。第一步，细胞在含有胰岛素、表皮生长因子（EGF）和亚油酸的培养基中的胶原凝胶内生长9天；这些条件下酪蛋白积累极少。在这个生长阶段之后，将凝胶释放使其漂浮在含有胰岛素、催乳素和亚油酸的培养基中。在这个第二阶段，乳腺细胞会积累大量酪蛋白，但前提是胰岛素、催乳素和亚油酸同时存在；在没有亚油酸的情况下，酪蛋白积累会大大减少。酪蛋白积累不依赖于糖皮质激素皮质醇的存在，并且即使存在抗糖皮质激素药物RU 38 486也会发生。为了确定其他研究者在器官培养中观察到的对皮质醇的反应是否可能由基质细胞介导，上皮细胞在无脂肪酸条件下的胶原凝胶中生长，然后与来自成年处女小鼠的乳腺脂肪垫外植体（有或没有乳腺实质）共培养。共培养在含有胰岛素和催乳素且有或没有皮质醇的无脂肪酸培养基中进行。在大多数实验中，无论外植体中是否存在乳腺实质，胶原凝胶中的乳腺上皮细胞在有皮质醇存在时比没有皮质醇时积累更多的酪蛋白。因此，乳腺上皮细胞直接依赖胰岛素和催乳素进行酪蛋白积累，并且通过其对基质细胞的作用间接依赖皮质醇。这种皮质醇效应可能是导致亚油酸或亚油酸的代谢产物从基质细胞释放到培养基中。
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