Newbold R R, Hanson R B, Jefferson W N
Developmental Endocrinology and Pharmacology Section, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina 27709.
In Vitro Cell Dev Biol Anim. 1994 Aug;30A(8):519-28.
Although estrogens have been shown to stimulate a variety of morphologic and biochemical changes in the uterus in vivo, no clear consistent demonstration of similar responses in vitro have been made; thus, a defined organ culture system using the immature mouse uterus was established to study the possibility of demonstrating estrogenic responses in vitro. Uterine tissue from immature outbred mice (17 to 24 days of age) were cut crosswise in 1-mm3 coins and cultured in a defined medium in the absence of serum, phenol red, or growth factor supplements. Diethylstilbestrol (DES), a synthetic estrogen, was added to the media at doses ranging from 1 to 100 ng/ml. The effect of DES on uterine cell proliferation was assessed by morphologic changes in uterine epithelial and stromal cells, increase in number of epithelial cells per unit basement membrane, increase in height of luminal epithelial cells, and [3H]thymidine incorporation. Functional changes were determined by measuring the amounts of the estrogen-inducible uterine protein, lactoferrin, that was localized in the epithelial cells and secreted into the media, and the localization of the estrogen receptor in the cultured tissues. Results indicate that under the described conditions of culture, estrogens like DES can induce morphologic and biochemical responses in the uterus that are similar to those seen in vivo. This organ culture system will aid in the investigation of various mechanisms involved in the hormonal regulation of growth and differentiation of estrogen target tissues.
尽管雌激素已被证明在体内能刺激子宫发生多种形态和生化变化，但在体外尚未明确一致地证明有类似反应；因此，建立了一种使用未成熟小鼠子宫的特定器官培养系统，以研究在体外证明雌激素反应的可能性。将未成熟远交系小鼠（17至24日龄）的子宫组织横向切成1立方毫米的薄片，在无血清、酚红或生长因子补充剂的特定培养基中培养。向培养基中添加合成雌激素己烯雌酚（DES），剂量范围为1至100纳克/毫升。通过子宫上皮和基质细胞的形态变化、每单位基底膜上皮细胞数量的增加、腔面上皮细胞高度的增加以及[3H]胸腺嘧啶核苷掺入来评估DES对子宫细胞增殖的影响。通过测量雌激素诱导的子宫蛋白乳铁蛋白的量来确定功能变化，乳铁蛋白定位于上皮细胞并分泌到培养基中，同时确定雌激素受体在培养组织中的定位。结果表明，在所描述的培养条件下，像DES这样的雌激素能在子宫中诱导出与体内所见类似的形态和生化反应。这种器官培养系统将有助于研究雌激素靶组织生长和分化的激素调节所涉及的各种机制。
