Luciferase transduction and selection protocol for reliable in vivo bioluminescent measurements in cancer research

  • Heliyon. 2024 Jun 23;10(13):e33356. doi: 10.1016/j.heliyon.2024.e33356.
Natacha Dehaen  1  2 ,  Matthias Van Hul  2  3 ,  Lionel Mignion  1  4 ,  Axell-Natalie Kouakou  1  2  5 ,  Patrice D Cani  2  3  6 ,  Bénédicte F Jordan  1  4
Affiliations
  • 1. Biomedical Magnetic Resonance group (REMA), Louvain Drug Research Institute (LDRI), UCLouvain, Université catholique de Louvain, Brussels, Belgium.
  • 2. Metabolism and Nutrition Research Group (MNUT), Louvain Drug Research Institute (LDRI), UCLouvain, Université catholique de Louvain, Brussels, Belgium.
  • 3. Walloon Excellence in Life Sciences and BIOtechnology (WELBIO), WELBIO department, WEL Research Institute, Wavre, Belgium.
  • 4. Nuclear and Electron Spin Technologies (NEST) Platform, Louvain Drug Research Institute (LDRI), UCLouvain, Université catholique de Louvain, Brussels, Belgium.
  • 5. Bioanalysis and Pharmacology of Bioactive Lipids (BPBL), Louvain Drug Research Institute (LDRI), UCLouvain, Université catholique de Louvain, Brussels, Belgium.
  • 6. Institute of Experimental and Clinical Research (IREC), UCLouvain, Université catholique de Louvain, Brussels, Belgium.
PMID: 39035528 DOI: 10.1016/j.heliyon.2024.e33356
Abstract

Bioluminescence imaging has become an essential non-invasive tool in Cancer research for monitoring various cellular processes and tumor progression in vivo. In this article, we aimed to propose a transduction and selection protocol for reliable in vivo bioluminescent measurements in immunocompetent mouse models. Using two different heterogenous luciferase-expressing cell models, we underlined factors influencing transduction. The protocol was tested through an in vitro luciferase activity assay as well as using in vivo longitudinal monitoring of metastases formation (In Vivo Imaging System®). The data were cross validated with histological assessment. Our results demonstrated stable and proportional in vitro and in vivo bioluminescent signals correlating with actual metastatic burden. Furthermore, ex vivo analysis confirmed the accuracy of bioluminescent imaging in quantifying metastatic surface area. This protocol should ensure reliable and reproducible measurements in Cancer research utilizing luciferase-positive cell lines, confirming the validity and accuracy of preclinical studies in immunocompetent models.

Keywords
Bioluminescence imaging; In vivo monitoring of metastasis; Luciferase; Transduction.