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Engineering small tubes with changes in diameter for the study of kidney cell organization

机译:工程直径变化的小管用于研究肾细胞的组织

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摘要

Multicellular tubes are structures ubiquitously found during development and in adult organisms. Their topologies (diameter, direction or branching), together with their mechanical characteristics, play fundamental roles in organ function and in the emergence of pathologies. In tubes of micrometric range diameters, typically found in the vascular system, renal tubules or excretory ducts, cells are submitted to a strong curvature and confinement effects in addition to flow. Then, small tubes with change in diameter are submitted to a local gradient of shear stress and curvature, which may lead to complex mechanotransduction responses along tubes, and may be involved in the onset or propagation of cystic or obstructive pathologies. We describe here a simple method to build a microfluidic device that integrates cylindrical channels with changes in diameter that mimic in vivo tube geometries. This microfabrication approach is based on molding of etched tungsten wires, which can achieve on a flexible way any change in diameter in a polydimethylsiloxane (PDMS) microdevice. The interest of this biomimetic multitube system has been evidenced by reproducing renal tubules on chip. In particular, renal cell lines were successfully seeded and grown in PDMS circular tubes with a transition between 80 μm and 50 μm diameters. Thanks to this biomimetic platform, the effect of the tube curvature has been investigated especially regarding cell morphology and orientation. The effect of shear stress on confluent cells has also been assessed simultaneously in both parts of tubes. It is thus possible to study interconnected cell response to differential constraints which is of central importance when mimicking tubes present in the organism.
机译:多细胞管是在发育过程中和成年生物中普遍发现的结构。它们的拓扑结构(直径,方向或分支)及其机械特性在器官功能和病理学出现中起着基本作用。在通常在血管系统,肾小管或排泄管中发现的,具有微米范围直径的管中,细胞除了具有流动性外,还具有很强的曲率和约束作用。然后,直径变化的小管会承受剪切应力和曲率的局部梯度,这可能会导致沿管的复杂的机械传导反应,并可能参与囊性或阻塞性疾病的发作或传播。我们在这里描述一种简单的方法来构建微流控设备,该设备集成了具有模拟体内管几何形状的直径变化的圆柱形通道。这种微细加工方法基于模制的蚀刻钨丝,可以灵活地实现聚二甲基硅氧烷(PDMS)微器件中直径的任何变化。这种仿生多管系统的兴趣已通过在芯片上繁殖肾小管得到了证明。尤其是,肾细胞系已成功播种并在直径为80μm至50μm的PDMS圆管中生长。由于这种仿生平台,已经研究了管曲率的影响,特别是在细胞形态和方向方面。还同时在试管的两个部分评估了剪应力对融合细胞的影响。因此,有可能研究相互连接的细胞对不同限制的反应,这在模仿生物体中存在的试管时至关重要。

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