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

• Published in: Analytical biochemistry Vol. 650; p. 114723
• Main Authors: Jessen, Stine Bull, Özkul, Derya Coskun, Özen, Yasemin, Gögenur, Ismail, Troelsen, Jesper T.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.08.2022
• Subjects: Cancer cell lines; Cell adhesion; Cell proliferation; Cellular assay; CRISPR; Luciferase; Cell proliferation; PBS; CRISPR; PS; Luciferase; CI; RTCA; Cancer cell lines; DMEM; CRC; FBS; Cell adhesion; Cellular assay; RLU
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2022.114723

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. 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. 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. 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. [Display omitted] •Stable luciferase-secreting cancer cell lines were generated using CRISPR.•Luciferase based assays were validated using RTCA iCELLigence impedance assay.•Luciferase proliferation and adhesion assays are highly reliable and sensitive.•These novel high-throughput assays overcome the limitations of existing methods.