Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system

• Main Authors: Teow, Sin Yeang *, Liew, Kitson, Che Mat, M., Marini, M., Norazlin, A. A., Chu, Tai Lin, Munirah, A., Khoo, Alan Soo Beng
• Format: Article
• Language: English
• Published: BMC (Springer Nature) 2019
• Subjects: QH301 Biology; R Medicine (General)
• Online Access: http://eprints.sunway.edu.my/1152/1/Teow%20Sin%20Yeang%20Development%20of%20a%20luciferaseluciferin%20cell.pdf; http://eprints.sunway.edu.my/1152/; http://doi.org/10.1186/s12896-019-0528-4

Background: In vitro modelling of cancer cells is becoming more complex due to prevailing evidence of intimate interactions between cancer cells and their surrounding stroma. A co-culture system which consists of more than one cell type is physiologically more relevant and thus, could serve as a useful model for various biological studies. An assay that specifically detects the phenotypic changes of cancer cells in a multi-cellular system is lacking for nasopharyngeal carcinoma (NPC). Results: Here, we describe a luciferase/luciferin (XenoLuc) assay that could specifically measure changes in the proliferation of cancer cells in the co-culture system using two modified NPC patient-derived tumour xenograft (PDTXs) cells: Xeno284-gfp-luc2 and XenoB110-gfp-luc2. Through this assay, we are able to show that the growth of NPC xenograft cells in both two-dimensional (2D) and three-dimensional (3D) models was enhanced when co-cultured with normal human dermal fibroblasts (NHDFs). In addition, potential applications of this assay in in vitro drug or inhibitor screening experiments are also illustrated. Conclusions: XenoLuc assay is specific, sensitive, rapid and cost-effective for measuring the growth of luciferase-expressing cells in a co- or multiple-culture system. This assay may also be adapted for tumour microenvironment studies as well as drug screening experiments in more complex 3D co-culture systems.