Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization.
Isolation of Mouse Endometrial Epithelial and Stromal Cells for In Vitro Decidualization.
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DOI：
10.3791/55168
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发表时间：
2017-03-02
期刊：
影响因子：
--
通讯作者：
Vriens J
中科院分区：
文献类型：
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作者：
De Clercq K;Hennes A;Vriens J
脱个体化是子宫内膜基质细胞依赖黄体酮的分化过程，是胚胎着床成功的先决条件。尽管许多研究已经揭示了脱个体化的潜在机制，但是与胚胎接触的上皮细胞和潜在的基质细胞之间的确切信号仍然知之甚少。因此，以兼顾上皮细胞和基质细胞的方式研究脱个体化可以提高我们对脱个体化分子细节的认识。为此，人工脱个体化的体内模型在生理学上是最相关的；然而，细胞间通讯的操纵是有限的。目前，子宫内膜基质细胞的体外培养正被用于研究几种信号分子对脱个体化的调节。通常，使用人类或小鼠子宫内膜基质细胞。然而，人类样本的可用性通常是有限的。此外，小鼠组织的使用伴随着培养方法的多样化。本研究提出了一种经过验证和标准化的方法，使用连续三天雌激素处理的成年完整小鼠获得纯子宫内膜上皮细胞（EEC）和基质细胞（ESC）培养物。优化方案以提高细胞的产量、活力和纯度，并进一步扩展以研究EEC和ESC共培养中的去个体化。该模型可能适合于利用这两种细胞类型在去个性化中的重要性，并评估在细胞间通讯中由EEC或ESC分泌的重要信号分子的贡献。
Decidualization is a progesterone-dependent differentiation process of endometrial stromal cells and is a prerequisite for successful embryo implantation. Although many efforts have been made to reveal the underlying mechanisms of decidualization, the exact signaling between the epithelial cells that are in contact with the embryo and the underlying stromal cells remains poorly understood. Therefore, studying decidualization in a way that takes both the epithelial and stromal cells into account could improve our knowledge about the molecular details of decidualization. For this purpose, in vivo models of artificial decidualization are physiologically the most relevant; however, manipulation of intercellular communication is limited. Currently, in vitro cultures of endometrial stromal cells are being used to investigate the modulation of decidualization by several signaling molecules. Conventionally, human or mouse endometrial stromal cells are used. However, the availability of human samples is very often limited. Furthermore, the use of murine tissues is accompanied with variety in the method of culturing. This study presents a validated and standardized method to obtain pure Endometrial Epithelial Cell (EEC) and Stromal Cell (ESC) cultures using adult intact mice treated with estrogen for three consecutive days. The protocol is optimized to improve the yield, viability, and purity of the cells and was further extended in order to study decidualization in a coculture of EEC and ESC. This model may be suitable to exploit the importance of both cell types in decidualization and to evaluate the contribution of significant signaling molecules secreted by EEC or ESC during the intercellular communication.