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

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 use...

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Autores principales: Teow, Sin-Yeang, Liew, Kitson, Che Mat, Mohd Firdaus, Marzuki, Marini, Abdul Aziz, Norazlin, Chu, Tai-Lin, Ahmad, Munirah, Khoo, Alan Soo-Beng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570829/
https://www.ncbi.nlm.nih.gov/pubmed/31200673
http://dx.doi.org/10.1186/s12896-019-0528-4
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author Teow, Sin-Yeang
Liew, Kitson
Che Mat, Mohd Firdaus
Marzuki, Marini
Abdul Aziz, Norazlin
Chu, Tai-Lin
Ahmad, Munirah
Khoo, Alan Soo-Beng
author_facet Teow, Sin-Yeang
Liew, Kitson
Che Mat, Mohd Firdaus
Marzuki, Marini
Abdul Aziz, Norazlin
Chu, Tai-Lin
Ahmad, Munirah
Khoo, Alan Soo-Beng
author_sort Teow, Sin-Yeang
collection PubMed
description 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12896-019-0528-4) contains supplementary material, which is available to authorized users.
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spelling pubmed-65708292019-06-27 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 Liew, Kitson Che Mat, Mohd Firdaus Marzuki, Marini Abdul Aziz, Norazlin Chu, Tai-Lin Ahmad, Munirah Khoo, Alan Soo-Beng BMC Biotechnol Methodology Article 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12896-019-0528-4) contains supplementary material, which is available to authorized users. BioMed Central 2019-06-14 /pmc/articles/PMC6570829/ /pubmed/31200673 http://dx.doi.org/10.1186/s12896-019-0528-4 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Teow, Sin-Yeang
Liew, Kitson
Che Mat, Mohd Firdaus
Marzuki, Marini
Abdul Aziz, Norazlin
Chu, Tai-Lin
Ahmad, Munirah
Khoo, Alan Soo-Beng
Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title_full Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title_fullStr Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title_full_unstemmed Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title_short Development of a luciferase/luciferin cell proliferation (XenoLuc) assay for real-time measurements of Gfp-Luc2-modified cells in a co-culture system
title_sort development of a luciferase/luciferin cell proliferation (xenoluc) assay for real-time measurements of gfp-luc2-modified cells in a co-culture system
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6570829/
https://www.ncbi.nlm.nih.gov/pubmed/31200673
http://dx.doi.org/10.1186/s12896-019-0528-4
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