A microfluidic bioreactor with integrated transepithelial electrical resistance (TEER) measurement electrodes for evaluation of renal epithelial cells.
A microfluidic bioreactor with integrated transepithelial electrical resistance (TEER) measurement electrodes for evaluation of renal epithelial cells.
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DOI：
10.1002/bit.22835
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发表时间：
2010-11-01
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;Desai, Ravi R.;Fleischman, Aaron J.;Roy, Shuvo;Humes, H. David;Fissell, William H.
我们开发了一种双层微流体系统，具有集成的跨上皮电阻（TEER）测量电极，用于在生理相关的流体流动条件下评估肾上皮细胞。该生物反应器由通过透明微孔膜连接的顶端和基底外侧流体室组成。顶部室包含微流体通道，用于灌注细胞的顶端表面。底部室充当跨细胞层运输的储存器，并为膜提供支撑。 TEER 电极集成到设备中，用于监测细胞生长并评估细胞间紧密连接的完整性。使用人肾上皮细胞 (HREC) 和 MDCK 细胞在微通道内对 ZO-1 紧密连接蛋白和乙酰化 α-微管蛋白（初级纤毛）进行免疫荧光染色。 HREC 对细胞骨架 F-肌动蛋白进行染色，并在暴露于剪切应力时表现出胞质 F-肌动蛋白应力纤维的解体。在正常培养条件下以及使用低 Ca2+ 培养基破坏紧密连接后，随时间监测 TEER。在 Ca2+ 转换之前和之后测量荧光标记示踪分子（FITC-菊粉）的转运速率，TEER 的降低与细胞旁菊粉转运的大幅增加相对应。这种生物反应器设计提供了一个具有生理意义的流动条件的仪器化平台，用于研究各种上皮细胞转运过程。
We have developed a bilayer microfluidic system with integrated transepithelial electrical resistance (TEER) measurement electrodes to evaluate kidney epithelial cells under physiologically relevant fluid flow conditions. The bioreactor consists of apical and basolateral fluidic chambers connected via a transparent microporous membrane. The top chamber contains microfluidic channels to perfuse the apical surface of the cells. The bottom chamber acts as a reservoir for transport across the cell layer and provides support for the membrane. TEER electrodes were integrated into the device to monitor cell growth and evaluate cell–cell tight junction integrity. Immunofluorescence staining was performed within the microchannels for ZO-1 tight junction protein and acetylated α-tubulin (primary cilia) using human renal epithelial cells (HREC) and MDCK cells. HREC were stained for cytoskeletal F-actin and exhibited disassembly of cytosolic F-actin stress fibers when exposed to shear stress. TEER was monitored over time under normal culture conditions and after disruption of the tight junctions using low Ca2+ medium. The transport rate of a fluorescently labeled tracer molecule (FITC-inulin) was measured before and after Ca2+ switch and a decrease in TEER corresponded with a large increase in paracellular inulin transport. This bioreactor design provides an instrumented platform with physiologically meaningful flow conditions to study various epithelial cell transport 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
DOI：
10.1088/0960-1317/18/6/067001
发表时间：
2008-06-01
影响因子：
2.3
作者：
Eddings, Mark A.;Johnson, Michael A.;Gale, Bruce K.
通讯作者：
Gale, Bruce K.
影响因子：
3.7
作者：
CHO, MJ;THOMPSON, DP;SCIESZKA, JF
通讯作者：
SCIESZKA, JF
影响因子：
6.1
作者：
Jang, Kyung-Jin;Suh, Kahp-Yang
通讯作者：
Suh, Kahp-Yang
影响因子：
4.2
作者：
Fissell, WH;Manley, S;Roy, S
通讯作者：
Roy, S