胎膜(羊膜绒毛膜),宫腔内的最内层,包围胎儿并包裹羊水。与单向血流不同,羊水微妙地来回摇摆,因此,最内层的羊膜上皮细胞持续暴露于流体波动引起的低水平剪切应力。这里,我们测试了液体运动对羊膜上皮细胞(AECs)的影响,作为一种力的影响,以及它们对可能破坏胎膜功能的细胞病理学变化的潜在脆弱性.使用了先前开发的羊膜(AM)芯片上器官(OOC),但具有动态流动来培养人胎儿羊膜细胞。调节施加的流量以来回灌注培养基48小时以模拟流体运动。静态培养条件用作阴性对照,氧化应激(OS)条件用作代表病理生理变化的阳性对照。通过测量细胞活力来评估流体运动的影响,细胞过渡,和炎症。此外,进行扫描电子显微镜(SEM)成像以观察微绒毛形成。结果表明,无论应用流量如何,AEC和AMC保持了它们的生存能力,形态学,先天元状态,和低产生的促炎细胞因子。E-cadherin表达和AECs中的微绒毛形成以流速依赖性方式上调;然而,这并不影响细胞形态或细胞转化或炎症.OS治疗诱导间质形态,波形蛋白与细胞角蛋白18(CK-18)的比率显着提高,和AECs中促炎细胞因子的产生,而AMC没有任何显著的反应。流体运动和剪切应力,如果有的话,不影响AEC细胞功能,也不引起炎症。因此,当使用羊膜OOC模型时,包含动态流动环境对于模拟子宫内羊膜的生理细胞条件是不必要的。
Fetal membrane (amniochorion), the innermost lining of the intrauterine cavity, surround the fetus and enclose amniotic fluid. Unlike unidirectional blood flow, amniotic fluid subtly rocks back and forth, and thus, the innermost amnion epithelial cells are continuously exposed to low levels of shear stress from fluid undulation. Here, we tested the impact of fluid motion on amnion epithelial cells (AECs) as a bearer of force impact and their potential vulnerability to cytopathologic changes that can destabilize fetal membrane functions. A previously developed amnion membrane (AM) organ-on-chip (OOC) was utilized but with dynamic flow to culture human fetal amnion membrane cells. The applied flow was modulated to perfuse culture media back and forth for 48 h to mimic fluid motion. A static culture condition was used as a negative control, and oxidative stress (OS) condition was used as a positive control representing pathophysiological changes. The impacts of fluidic motion were evaluated by measuring cell viability, cellular transition, and inflammation. Additionally, scanning electron microscopy (SEM) imaging was performed to observe microvilli formation. The results show that regardless of the applied flow rate, AECs and AMCs maintained their viability, morphology, innate meta-state, and low production of pro-inflammatory cytokines. E-cadherin expression and microvilli formation in the AECs were upregulated in a flow rate-dependent fashion; however, this did not impact cellular morphology or cellular transition or inflammation. OS treatment induced a mesenchymal morphology, significantly higher vimentin to cytokeratin 18 (CK-18) ratio, and pro-inflammatory cytokine production in AECs, whereas AMCs did not respond in any significant manner. Fluid motion and shear stress, if any, did not impact AEC cell function and did not cause inflammation. Thus, when using an amnion membrane OOC model, the inclusion of a dynamic flow environment is not necessary to mimic in utero physiologic cellular conditions of an amnion membrane.