Imaging- and flow cytometry-based analysis of cell position and the cell cycle in 3d melanoma spheroids

Beaumont, Kimberley A., Anfosso, Andrea, Ahmed, Farzana, Weninger, Wolfgang and Haass, Nikolas K. (2015) Imaging- and flow cytometry-based analysis of cell position and the cell cycle in 3d melanoma spheroids. Journal of Visualized Experiments, 106 e53486: . doi:10.3791/53486


Author Beaumont, Kimberley A.
Anfosso, Andrea
Ahmed, Farzana
Weninger, Wolfgang
Haass, Nikolas K.
Title Imaging- and flow cytometry-based analysis of cell position and the cell cycle in 3d melanoma spheroids
Journal name Journal of Visualized Experiments   Check publisher's open access policy
ISSN 1940-087X
Publication date 2015-12-28
Year available 2015
Sub-type Article (original research)
DOI 10.3791/53486
Open Access Status Not yet assessed
Volume 106
Issue e53486
Total pages 9
Publisher Journal of Visualized Experiments
Collection year 2016
Language eng
Abstract Three-dimensional (3D) tumor spheroids are utilized in cancer research as a more accurate model of the in vivo tumor microenvironment, compared to traditional two-dimensional (2D) cell culture. The spheroid model is able to mimic the effects of cell-cell interaction, hypoxia and nutrient deprivation, and drug penetration. One characteristic of this model is the development of a necrotic core, surrounded by a ring of G1 arrested cells, with proliferating cells on the outer layers of the spheroid. Of interest in the cancer field is how different regions of the spheroid respond to drug therapies as well as genetic or environmental manipulation. We describe here the use of the fluorescence ubiquitination cell cycle indicator (FUCCI) system along with cytometry and image analysis using commercial software to characterize the cell cycle status of cells with respect to their position inside melanoma spheroids. These methods may be used to track changes in cell cycle status, gene/protein expression or cell viability in different sub-regions of tumor spheroids over time and under different conditions.
Keyword Medicine
Melanoma
Spheroid
Cell cycle
Image analysis
Flow cytometry analysis
3D
Cancer therapy
Cancel cell biology
Hypoxia
Tumor sub-populations
Vibratome sectioning
Multicellular
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2016 Collection
UQ Diamantina Institute Publications
 
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Created: Mon, 18 Jan 2016, 17:33:38 EST by Nikolas Haass on behalf of Learning and Research Services (UQ Library)