A whole-cell biosensor as in vitro alternative to skin irritation tests.
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 μSlidesI(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.
Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Carl-Zeiss-Promenade 2, 07745 Jena, Germany.
Ute
U
Hofmann
Stefanie
S
Michaelis
Thomas
T
Winckler
Joachim
J
Wegener
Karl-Heinz
KH
Feller
D015374:Biosensing Techniques / Q000295:instrumentation*; D019256:Cadmium Chloride / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D017097:Electric Impedance; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D050884:HSP72 Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D056656:Lab-On-A-Chip Devices; D023422:Skin Irritancy Tests / Q000295:instrumentation*; D014162:Transfection
D050884:HSP72 Heat-Shock Proteins; D049452:Green Fluorescent Proteins; D019256:Cadmium Chloride