期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 106, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/53486
关键词
Medicine; Issue 106; Melanoma; spheroid; cell cycle; image analysis; flow cytometry analysis; 3D; cancer therapy; cancer cell biology; hypoxia; tumor sub-populations; vibratome sectioning; multicellular
资金
- Cancer Council New South Wales [RG 09-08, RG 13-06]
- Priority-driven collaborative cancer research scheme/Cancer Australia/Cure Cancer Australia Foundation [570778, 1051996]
- Cancer Institute New South Wales [08/RFG/1-27]
- National Health and Medical Research Council [APP1003637, APP1084893]
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.
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