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

Teow, Sin Yeang * and Liew, Kitson and Che Mat, M. and Marini, M. and Norazlin, A. A. and Chu, Tai Lin and Munirah, A. and Khoo, Alan Soo Beng (2019) Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system. BMC Biotechnology, 19 (34). ISSN 1472-6750

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Official URL: http://doi.org/10.1186/s12896-019-0528-4

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.

Item Type: Article
Uncontrolled Keywords: Luciferase assay; Luminescence; Nasopharyngeal carcinoma; Nasopharyngeal neoplasms; Patient-derived xenograft; Fibroblasts; Tumour microenvironment; Stromal interaction; Proliferation; Co-culture; Spheroids; Drug screening; Cancer immunology
Subjects: Q Science > QH Natural history > QH301 Biology
R Medicine > R Medicine (General)
Divisions: Others > Non Sunway Academics
Sunway University > School of Medical and Life Sciences [formerly School of Healthcare and Medical Sciences until 2020] > Department of Medical Sciences
Depositing User: Dr Janaki Sinnasamy
Related URLs:
Date Deposited: 28 Nov 2019 03:59
Last Modified: 28 Nov 2019 03:59
URI: http://eprints.sunway.edu.my/id/eprint/1152

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