Three Dimensional Modeling of Biologically Relevant Fluid Shear Stress in Human Renal Proximal Tubule Cells Mimics In Vivo Transcriptional Profiles. Three Dimensional Modeling of Biologically Relevant Fluid Shear Stress in Human Renal Proximal Tubule Cells Mimics In Vivo Transcriptional Profiles

We performed RNA-sequencing of human RPTEC/TERT1 cells in a microfluidic chip-based 3D model to determine transcriptomic changes. We measured transcriptional changes following the treatment of cells in this device at three different fluidic shear stress. We observed that FSS changes the expression of proximal tubule cell (PTC)-specific genes and impacted genes previously associated with renal diseases in genome-wide association studies (GWAS). At a physiological FSS level, we observed cell morphology, enhanced polarization, presence of cilia, and transport functions using albumin reabsorption via endocytosis and efflux transport. Here, we present a dynamic view of human PTCs response to FSS with increasing fluidic shear stress conditions and provide insight into hPTCs cellular function under biologically relevant conditions. Overall design: Examination of cells placed under flow rates treatments of 0.1dyn/cm2, 0.25 dyn/cm2, and a physiological level of 0.5 dyn/cm2

本研究依托微流控芯片三维模型，对人RPTEC/TERT1细胞开展RNA测序，以分析其转录组变化。本研究在该微流控装置中对细胞施加三种不同水平的流体剪切应力（fluidic shear stress, FSS），并检测该处理后细胞的转录组变化。研究发现，FSS可调控近端小管细胞（proximal tubule cell, PTC）特异性基因的表达，并影响既往全基因组关联研究（genome-wide association studies, GWAS）中与肾脏疾病相关的基因。在生理水平FSS条件下，我们观察到细胞形态完整、极化水平提升、纤毛形成，以及通过内吞作用重吸收白蛋白和外排转运所体现的转运功能。本研究呈现了人PTC随流体剪切应力升高的动态应答特征，并为阐明生理相关条件下hPTC的细胞功能提供了新的见解。实验整体设计：将细胞分别置于0.1dyn/cm²、0.25dyn/cm²及生理水平0.5dyn/cm²的流体剪切应力条件下进行检测。