期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 104, 期 3, 页码 516-525出版社
WILEY
DOI: 10.1002/bit.22413
关键词
fluorescence optical detection; microfluidics; cell culture analog; P450 enzyme; liver cells
资金
- Nanobiotechnology center (NBTC) [CM-2]
- National Science Foundation (NSF)
- Cornell Nanoscale Science and Technology Facility (CNF)
- Army Corp of Engineers [W9132T-07]
- Ministry of Education, Science, and Technology [R15-2004-024-00000-0, KOSEF 2007-8-1158, R01-2007-000-20821-0]
- MEMS Research Center for National Defense
- Ministry of Knowledge Economy of Korea [10030573]
We describe an in situ fluorescence optical detection system to demonstrate real-time and non-invasive detection of reaction products in a microfluidic device while under perfusion within a standard incubator. The detection system is designed to be compact and robust for operation inside a mammalian cell culture incubator for quantitative detection of fluorescent signal from microfluidic devices. When compared to a standard plate reader, both systems showed similar biphasic response curves with two linear regions. Such a detection system allows real-time measurements in microfluidic devices with cells without perturbing the culture environment. In a proof-of-concept experiment, the cytochrome P450 1A1/1A2 activity of a hepatoma cell line (HepG2/C3A) was monitored by measuring the enzymatic conversion of ethoxyresorufin to resorufin. The hepatoma cell line was embedded in Matrigel (TM) construct and cultured in a microfluidic device with medium perfusion. The response of the cells, in terms of P450 1A1/1A2 activity, was significantly different in a plate well system and the microfluidic device. Uninduced cells showed almost no activity in the plate assay, while uninduced cells in Matrigel (TM) with perfusion in a microfluidic device showed high activity. Cells in the plate assay showed a significant response to induction with 3-Methylcholanthrene while cells in the microfluidic device did not respond to the inducer. These results demonstrate that the system is a potentially useful method to measure cell response in a microfluidic system. Biotechnol. Bioeng. 2009;104: 516-525. (C) 2009 Wiley Periodicals, Inc.
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