A cell-based sensor system for toxicity testing using multiwavelength fluorescence spectroscopy

被引:13
作者
Fritzsche, Michael [1 ]
Fredriksson, J. Magnus [1 ]
Carlsson, Maria [1 ]
Mandenius, Carl-Fredrik [1 ]
机构
[1] Linkoping Univ, Div Biotechnol, IFM, S-58183 Linkoping, Sweden
关键词
Autofluorescence; HL-1 cell line; Cardiomyocytes; Cytotoxicity screening; Drug testing; Microtiter plate; Tryptophan fluorescence; In vitro toxicity; SCREENING ASSAY; BIOSENSORS;
D O I
10.1016/j.ab.2009.01.027
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A novel cell-based fluorometric sensor system for toxicity monitoring is described, which uses functional spontaneously contracting cardiomyocytes (HL-1 cell line) as the biological recognition element. Based on these highly specialized cells, it has the potential of providing a sensitive and relevant analytical in vitro toxicity testing method. The system was configured by propagating the surface-attaching HL-1 cardiomyocytes in the wells of a 96-well microtiter plate and connecting the plate via an optical fiber to a fluorescence spectrometer capable of excitation-emission matrix scanning. The fluorescence data were analyzed using a conventional spectral analysis software program. The performance of the system for detection of general cytotoxicity to the cells was evaluated using three well-known drugs: verapamil, quinidine, and acetaminophen. The dose-response curves were assessed and the EC50 values were determined (0.10 +/- 0.007, 0.23 +/- 0.025, and 12.32 +/- 2.40 mM, respectively). Comparison with in vitro and in vivo reference data for the drugs showed good correlations, suggesting that this cell-based sensor system Could be a useful tool in pharmacological in vitro drug testing. (C) 2009 Elsevier Inc. All rights reserved.
引用
收藏
页码:271 / 275
页数:5
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