Establishment of a luciferase-based method for measuring cancer cell adhesion and proliferation

Stine Bull Jessen, Derya Coskun Özkul, Yasemin Özen, Ismail Gögenur, Jesper T Troelsen*

*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskriftArtikelForskningpeer review

Abstract

BACKGROUND: Methods measuring cell proliferation and adhesion are widely used but each hold limitations. We, therefore, introduce novel methods for measuring cell proliferation and adhesion based on CRISPR-modified cancer cell lines secreting luciferase to the growth media.

MATERIALS AND METHODS: Using CRISPR genome editing, we generated stable luciferase-secreting LS174T, HCT 116, Caco-2, and PANC-1 cell lines. The modified cells were seeded, and luciferase activity was measured in the media and compared to Coulter counter cell counts and iCELLigence impedance assay to evaluate the value of the secreted luciferase activities as a measurement for adhesion and proliferation.

RESULTS: Our results demonstrate that luciferase secreted into the media can be used quantifying cell proliferation and adhesion. The adhesion luciferase assay and the iCELLigence impedance assay showed similar results with increased significant difference observed in the luciferase assays. The luciferase proliferation assay showed increased growth following increased serum concentrations in all cell lines vs. only two cell lines in the iCELLigence impedance assay.

CONCLUSIONS: Our results show that the luciferase adhesion and proliferation assays are reliable methods for measuring adhesion and proliferation. The luciferase assays have advantages over existing assays as they are highly sensitive, easy to perform, non-invasive and suitable as high-throughput measurements.

OriginalsprogEngelsk
Artikelnummer114723
TidsskriftAnalytical Biochemistry
Vol/bind650
Tidlig onlinedato12 maj 2022
DOI
StatusUdgivet - 1 aug. 2022

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Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.

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