Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium.
Compartmentalized Culture of Perivascular Stroma and Endothelial Cells in a Microfluidic Model of the Human Endometrium.
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DOI：
10.1007/s10439-017-1797-5
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发表时间：
2017-07
影响因子：
3.8
通讯作者：
Osteen KG
中科院分区：
文献类型：
--
作者：
Gnecco JS;Pensabene V;Li DJ;Ding T;Hui EE;Bruner-Tran KL;Osteen KG
子宫内膜是子宫的内层。在特定的周期性激素刺激后，子宫内膜间质成纤维细胞（基质）和血管内皮细胞表现出形态和生化变化，以支持胚胎着床和调节血管功能，分别。本文中，我们将基于树脂的多孔膜集成在聚二甲基硅氧烷中的双室微流体装置中，其允许人子宫内膜基质细胞和内皮细胞的长期体外共培养。这种透明的2 μm多孔膜将两个腔室分开，允许小分子扩散，并实现高分辨率亮场和荧光成像。在我们的基质和内皮细胞的原代人类共培养模型中，我们模拟了理想化的28天月经周期期间发生的时间激素变化。我们观察到基质成功分化为功能性蜕膜细胞，通过形态学和生物化学测定，通过增加催乳素的产生来确定。通过控制装置的微流体性质，我们还发现剪切应力促进了内皮层中细胞骨架的排列和紧密连接的形成。最后，我们证明了子宫内膜血管周围间质模型可持续长达4周，对类固醇保持敏感，并适用于定量生化分析。该装置的未来利用将允许直接评估这两种细胞类型之间的旁分泌和内分泌串扰，以及与正常与疾病相关的子宫内膜微环境相关的免疫学事件的研究。本文的在线版本（doi：10.1007/s10439-017-1797-5）包含补充材料，可供授权用户使用。
The endometrium is the inner lining of the uterus. Following specific cyclic hormonal stimulation, endometrial stromal fibroblasts (stroma) and vascular endothelial cells exhibit morphological and biochemical changes to support embryo implantation and regulate vascular function, respectively. Herein, we integrated a resin-based porous membrane in a dual chamber microfluidic device in polydimethylsiloxane that allows long term in vitro co-culture of human endometrial stromal and endothelial cells. This transparent, 2-μm porous membrane separates the two chambers, allows for the diffusion of small molecules and enables high resolution bright field and fluorescent imaging. Within our primary human co-culture model of stromal and endothelial cells, we simulated the temporal hormone changes occurring during an idealized 28-day menstrual cycle. We observed the successful differentiation of stroma into functional decidual cells, determined by morphology as well as biochemically as measured by increased production of prolactin. By controlling the microfluidic properties of the device, we additionally found that shear stress forces promoted cytoskeleton alignment and tight junction formation in the endothelial layer. Finally, we demonstrated that the endometrial perivascular stroma model was sustainable for up to 4 weeks, remained sensitive to steroids and is suitable for quantitative biochemical analysis. Future utilization of this device will allow the direct evaluation of paracrine and endocrine crosstalk between these two cell types as well as studies of immunological events associated with normal vs. disease-related endometrial microenvironments. The online version of this article (doi:10.1007/s10439-017-1797-5) contains supplementary material, which is available to authorized users.