Albumin handling by renal tubular epithelial cells in a microfluidic bioreactor.
Albumin handling by renal tubular epithelial cells in a microfluidic bioreactor.
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DOI：
10.1002/bit.24339
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发表时间：
2012-03
影响因子：
3.8
通讯作者：
Fissell, William H.
中科院分区：
文献类型：
--
作者：
Ferrell, Nicholas;Ricci, Kevin B.;Groszek, Joseph;Marmerstein, Joseph T.;Fissell, William H.
肾近端小管上皮细胞从肾小球滤液中回收和代谢蛋白质。蛋白尿是尿中蛋白质过多，影响肾小管细胞功能，是慢性肾脏疾病进展的主要因素。通过开发实验系统来研究在模拟体内肾小管的一些环境条件的设置中的肾小管蛋白处理，我们可以更好地了解微环境条件如何影响细胞蛋白处理，以确定这些条件是否与疾病相关。为此，我们使用了两种体外微流体模型来评估肾近端小管细胞对白蛋白的处理。对于第一个系统，细胞在微流体通道中生长，并灌注生理水平的剪切应力，以评估机械应力对蛋白质摄取的作用。在第二个系统中，使用多孔膜分离顶侧和基底侧隔室，以评价细胞代谢后蛋白质的命运。将负鼠肾（OK）上皮细胞暴露于荧光标记的白蛋白，并通过测量细胞裂解物的荧光来确定细胞摄取。共聚焦荧光显微镜用于比较在流动和静态条件下生长的细胞中的摄取。通过尺寸排阻色谱（SEC）和SDS-PAGE检测细胞加工的白蛋白。结果显示，与静态条件相比，暴露于流动的细胞中的细胞摄取和/或降解显著增加。这通过共聚焦显微镜证实。大小排阻层析和SDS-PAGE显示，白蛋白被分解成小分子量片段并由细胞分泌。SEC或SDS-PAGE均未检测到痕量完整白蛋白。这些结果表明，流体剪切力是介导细胞蛋白质处理的重要因素，微流体体外模型提供了一种新的工具来研究这些过程。
Epithelial cells in the proximal tubule of the kidney reclaim and metabolize protein from the glomerular filtrate. Proteinuria, an overabundance of protein in the urine, affects tubular cell function and is a major factor in the progression of chronic kidney disease. By developing experimental systems to study tubular protein handling in a setting that simulates some of the environmental conditions of the kidney tubule in vivo, we can better understand how microenviromental conditions affect cellular protein handling to determine if these conditions are relevant in disease. To this end, we used two in vitro microfluidic models to evaluate albumin handling by renal proximal tubule cells. For the first system, cells were grown in a microfluidic channel and perfused with physiological levels of shear stress to evaluate the role of mechanical stress on protein uptake. In the second system, a porous membrane was used to separate an apical and basolateral compartment to evaluate the fate of protein following cellular metabolism. Opossum kidney (OK) epithelial cells were exposed to fluorescently labeled albumin, and cellular uptake was determined by measuring the fluorescence of cell lysates. Confocal fluorescence microscopy was used to compare uptake in cells grown under flow and static conditions. Albumin processed by the cells was examined by size exclusion chromatography (SEC) and SDS-PAGE. Results showed that cellular uptake and/or degradation was significantly increased in cells exposed to flow compared to static conditions. This was confirmed by confocal microscopy. Size exclusion chromatography and SDS-PAGE showed that albumin was broken down into small molecular weight fragments and excreted by the cells. No trace of intact albumin was detectable by either SEC or SDS-PAGE. These results indicate that fluid shear stress are important factors mediating cellular protein handling and microfluidic in vitro models provide a novel tool to investigate these processes.
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DOI：
10.1073/pnas.0804954105
发表时间：
2008-08-12
影响因子：
11.1
作者：
Duan, Yi;Gotoh, Nanami;Weinbaum, Sheldon
通讯作者：
Weinbaum, Sheldon
影响因子：
2.5
作者：
Clavant, SP;Greive, KA;Comper, WD
通讯作者：
Comper, WD
影响因子：
--
作者：
MAUNSBACH, AB;GIEBISCH, GH;STANTON, BA
通讯作者：
STANTON, BA
影响因子：
7.7
作者：
Osicka, TM;Houlihan, CA;Comper, WD
通讯作者：
Comper, WD
DOI：
10.1152/ajprenal.1995.268.5.f899
发表时间：
1995-05-01
期刊：
AMERICAN JOURNAL OF PHYSIOLOGY-RENAL FLUID AND ELECTROLYTE PHYSIOLOGY
影响因子：
--
作者：
GEKLE, M;MILDENBERGER, S;SILBERNAGL, S
通讯作者：
SILBERNAGL, S