A novel organotypic culture model for normal human endometrium:: regulation of epithelial cell proliferation by estradiol and medroxyprogesterone acetate
A novel organotypic culture model for normal human endometrium:: regulation of epithelial cell proliferation by estradiol and medroxyprogesterone acetate
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DOI：
10.1093/humrep/deh722
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发表时间：
2005-04-01
影响因子：
6.1
通讯作者：
Ylikomi, T
中科院分区：
文献类型：
--
作者：
Bläuer, M;Heinonen, PK;Ylikomi, T
背景：建立了一种新的器官型培养系统，用于模拟体外正常人子宫内膜的激素反应。方法：将子宫内膜上皮细胞作为腺样器官在组织培养插入物中的重组细胞外基质（Matrigel）中培养，将基质细胞在上皮室下面的塑料上培养。研究了雌二醇（E-2）和E-2与醋酸甲孕酮（MPA）对10例上皮-间质共培养和3例上皮类器官平行单培养细胞增殖及雌激素受体α (ER α)和孕激素受体（PR）表达的影响。结果：在共培养中，与未处理的对照组相比，E-2显示ki67阳性细胞的百分比增加了2倍。在MPA存在的情况下，细胞增殖明显减少。当计算ki67阳性类器官的相应百分比而不是单个细胞时，也得到了类似的结果。在缺乏间质成纤维细胞的情况下，两种激素治疗后Ki67上皮标记仍低于控制值。上皮类器官在培养中保留了表达雌激素和孕激素受体的能力。两种激素处理均能显著提高E-2的表达，而MPA可下调PR的表达，而ER α的表达仅受轻微影响。结论：目前的器官型模型提供了一个新的体外系统来研究类固醇对正常人子宫内膜细胞增殖和基因表达的影响。该培养系统有望成为筛选新型类固醇化合物的有用方法，并可能有助于规避与使用动物模型相关的问题。
BACKGROUND: A novel organotypic culture system was established for modelling the hormonal responses of the normal human endometrium in vitro. METHODS: Endometrial epithelial cells were cultured as glandular organoids within reconstituted extracellular matrix (Matrigel) in tissue culture inserts and stromal cells on plastic below the epithelial compartment. The effects of estradiol (E-2) and E-2 together with medroxyprogesterone acetate (MPA) on cell proliferation and the expression of estrogen receptor alpha (ER alpha) and progesterone receptor (PR) were studied in 10 epithelial-stromal co-cultures and in three parallell monocultures of epithelial organoids. RESULTS: In co-cultures, E-2 was shown to increase the percentage of Ki67-positive cells by similar to 2-fold relative to untreated controls. In the presence of MPA, a significant decrease in cell proliferation was detected. Similar results were obtained when the corresponding percentages of Ki67-positive organoids were calculated instead of individual cells. In the absence of stromal fibroblasts, Ki67 epithelial labelling remained below the control value after both hormonal treatments. Epithelial organoids retained their capacity to express estrogen and progesterone receptors in culture. E-2 was shown to markedly increase and MPA to down-regulate the expression of PR. The expression of ER alpha was only slightly affected by either hormonal treatment. CONCLUSIONS: The present organotypic model provides a novel in vitro system in which to study the effects of steroids in the normal human endometrium both in terms of cell proliferation and gene expression. The culture system holds promise as a useful method to screen novel steroid compounds and may help to circumvent problems related to the use of animal models.