1. Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system.
- Author
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Teow SY, Liew K, Che Mat MF, Marzuki M, Abdul Aziz N, Chu TL, Ahmad M, and Khoo AS
- Subjects
- Animals, Cell Line, Tumor, Cells, Cultured, Coculture Techniques, Fibroblasts cytology, Green Fluorescent Proteins genetics, HEK293 Cells, Humans, Luciferases genetics, Luminescent Measurements methods, Mice, Inbred NOD, Mice, Knockout, Mice, SCID, Nasopharyngeal Carcinoma genetics, Nasopharyngeal Carcinoma pathology, Transplantation, Heterologous, Fibroblasts metabolism, Green Fluorescent Proteins metabolism, Luciferases metabolism, Nasopharyngeal Carcinoma metabolism
- Abstract
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.
- Published
- 2019
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