Please use this identifier to cite or link to this item:https://hdl.handle.net/20.500.12259/52109
Type of publication: Straipsnis Clarivate Analytics Web of Science ar/ir Scopus / Article in Clarivate Analytics Web of Science or / and Scopus (S1)
Field of Science: Biofizika / Biophysics (N011)
Author(s): Jakštys, Baltramiejus;Ruzgys, Paulius;Tamošiūnas, Mindaugas;Šatkauskas, Saulius
Title: Different cell viability assays reveal inconsistent results after bleomycin electrotransfer in vitro
Is part of: Journal of membrane biology. New York : Springer-Verlag, Vol. 248, iss. 5, 2015
Extent: p. 857-863
Date: 2015
Note: ISSN 1432-1424 (Online)
Keywords: Elektroporacija;Vaistų pernaša;Elektropernaša;Ląstelių gyvybingumo testai;Tėkmės citometrija;Electroporation;Drug delivery;Electrotransfer;Bleomycin;Cell viability assays;Flow cytometry
Abstract: The aim of this study was to compare different and commonly used cell viability assays after CHO cells treatment with anticancer drug bleomycin (20 nM), high voltage (HV) electric pulses (4 pulses, 1200 V/cm, 100 µs, 1 Hz), and combination of bleomycin and HV electric pulses. Cell viability was measured using clonogenic assay, propidium iodide (PI) assay, MTT assay, and employing flow cytometry modality to precisely count cells in definite volume of the sample (flow cytometry assay). Results showed that although clonogenic cell viability drastically decreased correspondingly to 57 and 3 % after cell treatment either with HV pulses or combination of bleomycin and HV pulses (bleomycin electrotransfer), PI assay performed ~15 min after the treatments indicated nearly 100 % cell viability. MTT assay performed at 6–72 h time points after these treatments revealed that MTT cell viability is highly dependent on evaluation time point and decreased with later evaluation time points. Nevertheless, in comparison to clonogenic cell viability, MTT cell viability after bleomycin electrotransfer at all testing time points was significantly higher. Flow cytometry assay if used at later times, 2–3 days after the treatment, allowed reliable evaluation of cell viability. In overall, our results showed that in order to estimate cell viability after cell treatment with combination of the bleomycin and electroporation the most reliable method is clonogenic assay. Improper use of PI and MTT assays can lead to misinterpretation of the experimental results
Internet: http://link.springer.com/article/10.1007/s00232-015-9813-x/fulltext.html
http://link.springer.com/article/10.1007/s00232-015-9813-x/fulltext.html
Affiliation(s): Biochemijos katedra
Biologijos katedra
Gamtos mokslų fakultetas
Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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