The extracellular matrix (ECM) provides physical and chemical support to the surrounding cells. During cell growth, ECM secretion and network formation influence cell morphology, cell adhesion, cell-to-cell interactions, and cell migration. Microfluidics-based cell culture systems are limited by the integration of structural ECM into the device. We report the development of a cell-derived ECM-incorporated microfluidic device that can provide structural characteristics and biochemical components of cell-derived ECM. Using an on-chip decellularization process, we constructed an ECM sheet, secreted and deposited from monolayer-cultured mouse embryonic fibroblasts (NIH/3T3), inside the microfluidic device. ECM components (including collagens, fibronectin, laminin, and elastin) and mesh-type fibronectin fibrous architecture were maintained on the surface of the porous membrane of the microfluidic device after decellularization. To verify the usability of the fibroblast-derived ECM sheet integrated microfluidic device in a cell culture platform, we tested the recellularization of human umbilical vein endothelial cells (HUVEC) and analyzed HUVEC–ECM and HUVEC–HUVEC interactions. On the ECM sheet, HUVECs exhibited morphologies and focal adhesion features that were markedly different from those of other groups. We then explored the effect of the ECM sheet on HUVEC mechanosensitivity. An increase in fluid shear stresses led to focal adhesion and the polymerization and reorganization of HUVEC adherens junctions, similar to natural junctional development, whereas the control group exhibited stimuli-insensitive behaviors. We conclude that the decellularized ECM sheet-incorporated microfluidic device provides an in vivo-like physical and biochemical ECM microenvironment for microfluidics-based cell culture.

中文翻译：

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细胞来源的细胞外基质片的片上加工

细胞外基质（ECM）为周围的细胞提供物理和化学支持。在细胞生长期间，ECM分泌和网络形成会影响细胞形态，细胞粘附，细胞间相互作用和细胞迁移。基于微流体的细胞培养系统受到结构ECM集成到设备中的限制。我们报告了细胞衍生的ECM结合的微流控设备的发展，该设备可以提供细胞衍生的ECM的结构特征和生化成分。使用芯片上的脱细胞过程，我们在微流控设备内部构造了由单层培养的小鼠胚胎成纤维细胞（NIH / 3T3）分泌和沉积的ECM薄层。ECM成分（包括胶原蛋白，纤连蛋白，层粘连蛋白，和弹性蛋白）和网状纤连蛋白的纤维结构在脱细胞后保持在微流体装置的多孔膜的表面上。为了验证在细胞培养平台中源自成纤维细胞的ECM薄板集成微流控设备的可用性，我们测试了人脐静脉内皮细胞（HUVEC）的再细胞化作用，并分析了HUVEC–ECM和HUVEC–HUVEC的相互作用。在ECM纸上，HUVEC表现出的形态和粘着斑特征与其他组明显不同。然后，我们探讨了ECM薄板对HUVEC机械敏感性的影响。流体剪切应力的增加导致了黏着力以及HUVEC黏着连接的聚合和重组，这类似于自然的连接发展，对照组表现出对刺激不敏感的行为。我们得出的结论是，脱细胞的ECM板结合微流控设备为基于微流控的细胞培养提供了体内类似的物理和生化ECM微环境。