A whole-cell biosensor as in vitro alternative to skin irritation tests.
作者： Ute Hofmann , Stefanie Michaelis , Thomas Winckler , Joachim Wegener , Karl-Heinz Feller
DOI: 10.1016/J.BIOS.2012.07.075
关键词:
摘要: This study presents the time-resolved detection of chemically induced stress upon intracellular signaling cascades by using genetically modified sensor cells based on human keratinocyte cell line HaCaT. The were stably transfected with a HSP72-GFP reporter gene construct to create an optical expressing stress-inducible protein. time- and dose-dependent performance is demonstrated discussed in comparison label-free impedimetric monitoring approach (electric cell-substrate impedance sensing, ECIS). Moreover, microfluidic platform was established μSlidesI(0,4)Luer allow for convenient, sterile incubator-independent time-lapse microscopic observation cells. Cell growth successfully achieved this setup cellular response cytotoxic substance could be followed real-time non-invasive, sensitive manner. paves way development micro-total analysis systems that combine readouts enable overall quantitative characterization changes metabolism morphology as toxin exposure. By recording multiple parameters, detailed discrimination between competing stress- or growth-related mechanisms possible, thereby presenting entirely new vitro alternative skin irritation tests.
core.ac.uk
sci-hub.se 参考文章(26)
W D Morgan, G T Williams, R I Morimoto, J Greene, R E Kingston, R Tjian, Two transcriptional activators, CCAAT-box-binding transcription factor and heat shock transcription factor, interact with a human hsp70 gene promoter. Molecular and Cellular Biology. ,vol. 7, pp. 1129- 1138 ,(1987) , 10.1128/MCB.7.3.1129
Amy E. Palmer, Yan Qin, Jungwon Genevieve Park, Janet E. McCombs, Design and application of genetically encoded biosensors Trends in Biotechnology. ,vol. 29, pp. 144- 152 ,(2011) , 10.1016/J.TIBTECH.2010.12.004
R. I. Morimoto, Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators Genes & Development. ,vol. 12, pp. 3788- 3796 ,(1998) , 10.1101/GAD.12.24.3788
I.L.A. Boxman, P.J. Hensbergen, R.C. Van Der Schors, D.P. Bruynzeel, C.P. Tensen, M. Ponec, Proteomic analysis of skin irritation reveals the induction of HSP27 by sodium lauryl sulphate in human skin. British Journal of Dermatology. ,vol. 146, pp. 777- 785 ,(2002) , 10.1046/J.1365-2133.2002.04714.X
Judith A. Stolwijk, Christoph Hartmann, Poonam Balani, Silke Albermann, Charles R. Keese, Ivar Giaever, Joachim Wegener, Impedance analysis of adherent cells after in situ electroporation: Non-invasive monitoring during intracellular manipulations Biosensors and Bioelectronics. ,vol. 26, pp. 4720- 4727 ,(2011) , 10.1016/J.BIOS.2011.05.033
S Aıt-Aıssa, J-M Porcher, A-P Arrigo, C Lambre, None, Activation of the hsp70 promoter by environmental inorganic and organic chemicals: relationships with cytotoxicity and lipophilicity. Toxicology. ,vol. 145, pp. 147- 157 ,(2000) , 10.1016/S0300-483X(00)00145-1
Paul J. Hung, Philip J. Lee, Poorya Sabounchi, Robert Lin, Luke P. Lee, Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays. Biotechnology and Bioengineering. ,vol. 89, pp. 1- 8 ,(2005) , 10.1002/BIT.20289
Roger Emter, Graham Ellis, Andreas Natsch, Performance of a novel keratinocyte-based reporter cell line to screen skin sensitizers in vitro Toxicology and Applied Pharmacology. ,vol. 245, pp. 281- 290 ,(2010) , 10.1016/J.TAAP.2010.03.009
Hsiang-Yu Wang, Ning Bao, Chang Lu, A microfluidic cell array with individually addressable culture chambers Biosensors and Bioelectronics. ,vol. 24, pp. 613- 617 ,(2008) , 10.1016/J.BIOS.2008.06.005
Baoyue Zhang, Min-Cheol Kim, Todd Thorsen, Zhanhui Wang, A self-contained microfluidic cell culture system. Biomedical Microdevices. ,vol. 11, pp. 1233- 1237 ,(2009) , 10.1007/S10544-009-9342-4