Terry P M, Banerjee M R, Lui R M
Proc Natl Acad Sci U S A. 1977 Jun;74(6):2441-5. doi: 10.1073/pnas.74.6.2441.
The whole second thoracic mammary gland of estradiol-17beta + progesterone primed 3- to 4-week-old BALB/c female mice was induced to pregnancy-like lobulo-alveolar morphogenesis after 6-day cultivation in a serum-free culture medium containing a "growth promoting" hormone mixture, insulin + prolactin + growth hormone (somatotropin) + estradiol + progesterone. No radioimmunologically detectable casein was present in these glands. Subsequent cultivation for another 6 days in a "lactogenic" medium with the hormones insulin + prolactin + cortisol produced abundant milk-like secretory material in the alveolar lumen. RNA of the mammary gland after estradiol + progesterone priming or cultivation in the "growth-promoting" medium failed to show a measurable amount of casein mRNA activity when assayed in a cell-free protein synthesis system derived from Ehrlich ascites ribosomes, rabbit reticulocyte factors, and tRNA. However, the glands sequentially cultivated in the "growth-promoting" and the "lactogenic" media showed a high level of casein mRNA activity in the heterologous cell-free protein synthesis system. Sodium dodecyl sulfate/polyacrylamide gel electrophoretic characteristics of the immunoprecipitable (by antibody to mouse milk casein) polypeptides directed by the mammary RNA induced in organ culture medium containing the lactogenic hormones were similar to the characteristics of the polypeptides directed by mammary polysomes of lactating mice. These results demonstrate hormonal induction of a specific mRNA in a sequential two-step culture of an entire organ in a serum-free chemically defined medium.
将17β-雌二醇+孕酮预处理的3至4周龄BALB/c雌性小鼠的整个第二胸段乳腺，在含有“促生长”激素混合物（胰岛素+催乳素+生长激素（促生长素）+雌二醇+孕酮）的无血清培养基中培养6天后，诱导出类似妊娠的小叶-腺泡形态发生。这些腺体中不存在放射免疫可检测到的酪蛋白。随后在含有胰岛素+催乳素+皮质醇的“生乳”培养基中再培养6天，在腺泡腔中产生了大量类似乳汁的分泌物质。在由艾氏腹水核糖体、兔网织红细胞因子和tRNA组成的无细胞蛋白质合成系统中检测时，经雌二醇+孕酮预处理或在“促生长”培养基中培养后的乳腺RNA，未能显示出可测量的酪蛋白mRNA活性。然而，在“促生长”和“生乳”培养基中依次培养的腺体，在异源无细胞蛋白质合成系统中显示出高水平的酪蛋白mRNA活性。在含有生乳激素的器官培养基中诱导产生的乳腺RNA所指导的、可被（抗小鼠乳酪蛋白抗体）免疫沉淀的多肽的十二烷基硫酸钠/聚丙烯酰胺凝胶电泳特征，与泌乳小鼠乳腺多核糖体所指导的多肽特征相似。这些结果表明，在无血清化学限定培养基中对整个器官进行连续两步培养时，激素可诱导特定mRNA的产生。
