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Tetracycline-mediated fluorescence correlates with passage number and β-galactosidase activity in HeLa and HEK293T cells

Redwood-Sawyerr, Chileab; Nesbeth, Darren N; (2023) Tetracycline-mediated fluorescence correlates with passage number and β-galactosidase activity in HeLa and HEK293T cells. Biology Open , 12 (7) , Article bio059691. 10.1242/bio.059691. Green open access

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Abstract

We report data consistent with tetracycline-mediated fluorescence having the potential to be an effective marker of senescence in immortalised cells. HeLa cells that had previously undergone more than 20 passages were transiently transfected with a plasmid encoding a novel tetracycline-inducible transgene featuring an open reading frame for green fluorescent protein. While characterising the performance of this plasmid and transfection procedure, HeLa cell fluorescence was observed to result from incubating cells with media containing 2 μg/ml tetracycline alone, without plasmid or transfection reagent. To investigate this phenomenon further, HeLa and HEK293T cells were purchased from a tissue culture collection and after cultivation over 4-23 passages, incubated with media containing 2 μg/ml tetracycline. For both cell lines, tetracycline-mediated fluorescence increase correlated with passage number increase. This effect in HeLa and HEK293T cells was also borne out by expression of β-galactosidase activity, an imperfect but widely used marker of cellular senescence. These data suggest tetracycline may have utility as a marker of cellular senescence in immortal cells and can inform future investigation and validation of this previously unreported application for this reagent.

Type: Article
Title: Tetracycline-mediated fluorescence correlates with passage number and β-galactosidase activity in HeLa and HEK293T cells
Open access status: An open access version is available from UCL Discovery
DOI: 10.1242/bio.059691
Publisher version: https://doi.org/10.1242/bio.059691
Language: English
Additional information: Copyright © 2023. Published by The Company of Biologists Ltd This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
Keywords: Senescence, Tetracycline, Marker, Cellular, HeLa, HEK293T
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10170920
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