Fabrication of two-layered channel system with embedded electrodes to measure resistance across epithelial and endothelial barriers.
Fabrication of two-layered channel system with embedded electrodes to measure resistance across epithelial and endothelial barriers.
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DOI：
10.1021/ac9029345
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发表时间：
2010-03-15
影响因子：
7.4
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Douville, Nicholas J.;Tung, Yi-Chung;Li, Ran;Wang, Jack D.;El-Sayed, Mohamed E. H.;Takayama, Shuichi
本文描述了一种在双层PDMS微流控芯片中嵌入Ag/AgCl电极的简单制造工艺，其中上下通道由半多孔膜隔开。该系统允许使用阻抗谱对跨内皮细胞和跨上皮细胞的跨内皮和跨上皮电阻(TEER)进行可靠的实时测量，TEER是细胞单层完整性的公认量化指标。该技术消除了昂贵或专门的微电极制造的需要，使商业上可用的线电极能够轻松地整合到PDMS微系统中，用于在微流体环境下测量TEER。使用(I)脑源性内皮细胞(bEND.3)，(Ii)Madin Darby犬肾细胞(MDCK-2)和小鼠成肌细胞(C2C12)(均来自ATCC，Manassas，VA)证实了跨融合细胞单层测量阻抗的能力。TER值作为细胞类型和细胞培养时间的函数被测量，并且两者都与先前发表的来自大规模培养技术的值一致。该系统为方便地解决跨内皮和跨上皮电阻以实时监测微流控细胞培养中的细胞功能开辟了新的机会。
This manuscript describes a straightforward fabrication process for embedding Ag/AgCl electrodes within a two-layer PDMS microfluidic chip where an upper and a lower channel are separated by a semi-porous membrane. This system allows for the reliable real-time measurement of trans-endothelial and trans-epithelial electrical resistance (TEER), an accepted quantification of cell monolayer integrity, across cells cultured on membranes inside the microchannels using impedance spectroscopy. The technique eliminates the need for costly or specialized microelectrode fabrication, enabling commercially available wire electrodes to easily be incorporated into PDMS microsystems for measuring TEER under microfluidic environments. The capability of measuring impedance across a confluent cell monolayer is confirmed using (i) brain-derived endothelial cells (bEND.3), (ii) Madin Darby Canine Kidney Cells (MDCK-2), and mouse myoblast (C2C12) (all from ATCC, Manassas, VA). TEER values as a function of cell type and cell culture time were measured and both agree with previously published values from macro-scale culture techniques. This system opens new opportunities for conveniently resolving both trans-endothelial and trans-epithelial electrical resistance to monitor cell function in real-time in microfluidic cell cultures.
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DOI：
10.1016/0165-022x(95)00011-x
发表时间：
1995-11-01
影响因子：
--
作者：
ERBEN, M;DECKER, S;GALLA, HJ
通讯作者：
GALLA, HJ
影响因子：
7.4
作者：
García, CD;Henry, CS
通讯作者：
Henry, CS
影响因子：
8.4
作者：
Iliescu, Ciprian;Poenar, Daniel P.;Loe, Felicia C.
通讯作者：
Loe, Felicia C.
影响因子：
4.5
作者：
KOTTRA, G;FROMTER, E
通讯作者：
FROMTER, E
影响因子：
4.2
作者：
Garcia, CD;Henry, CS
通讯作者：
Henry, CS