Hofmann, Ute and Michaelis, Stefanie and Winckler, Thomas and Wegener, Joachim and Feller, Karl-Heinz (2013) A whole-cell biosensor as in vitro alternative to skin irritation tests. BIOSENSORS & BIOELECTRONICS, 39 (1). pp. 156-162. ISSN 0956-5663,
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This study presents the time-resolved detection of chemically induced stress upon intracellular signaling cascades by using genetically modified sensor cells based on the human keratinocyte cell line HaCaT. The cells were stably transfected with a HSP72-GFP reporter gene construct to create an optical sensor cell line expressing a stress-inducible reporter protein. The time- and dose-dependent performance of the sensor cells is demonstrated and discussed in comparison to a label-free impedimetric monitoring approach (electric cell-substrate impedance sensing, ECIS). Moreover, a microfluidic platform was established based on mu SlidesI(0.4)Luer to allow for a convenient, sterile and incubator-independent time-lapse microscopic observation of the sensor cells. Cell growth was successfully achieved in this microfluidic setup and the cellular response to a cytotoxic substance could be followed in real-time and in a non-invasive, sensitive manner. This study paves the way for the development of micro-total analysis systems that combine optical and impedimetric readouts to enable an overall quantitative characterization of changes in cell metabolism and morphology as a response to toxin exposure. By recording multiple parameters, a detailed discrimination between competing stress- or growth-related mechanisms is possible, thereby presenting an entirely new in vitro alternative to skin irritation tests. (C) 2012 Elsevier B.V. All rights reserved.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HUMAN EPIDERMIS; CULTURE; PROMOTER; PERFUSION; PROTEINS; BINDING; MODELS; ASSAYS; ARRAY; Lab on a Chip; Cell-based assay; Whole-cell biosensor; Heat shock protein; Reporter gene; Time-lapse microscopy; Electric cell-substrate impedance sensing |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Bioanalytik und Biosensorik (Prof. Joachim Wegener) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 28 Apr 2020 07:26 |
| Last Modified: | 28 Apr 2020 07:26 |
| URI: | https://pred.uni-regensburg.de/id/eprint/17289 |
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