Abstract
Aberrant cell cycle progression is a hallmark of solid tumors; therefore, cell cycle analysis is an invaluable technique to study cancer cell biology. However, cell cycle progression has been most commonly assessed by methods that are limited to temporal snapshots or that lack spatial information. Here, we describe a technique that allows spatiotemporal real-time tracking of cell cycle progression of individual cells in a multicellular context. The power of this system lies in the use of 3D melanoma spheroids generated from melanoma cells engineered with the fluorescent ubiquitination-based cell cycle indicator (FUCCI). This technique allows us to gain further and more detailed insight into several relevant aspects of solid cancer cell biology, such as tumor growth, proliferation, invasion, and drug sensitivity.
*These authors contributed equally to this work.
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Acknowledgments
We thank Prof. Meenhard Herlyn, The Wistar Institute, Philadelphia, PA; Prof. Keiran Smalley, Moffitt Cancer Center, Tampa, FL; Prof. Wolfgang Weninger & Dr. Ben Roediger, Centenary Institute, Sydney, NSW; Dr. Crystal Tonnessen, UQ Diamantina Institute, Brisbane, QLD, and the imaging facilities of the Centenary and UQ Diamantina Institutes for their contribution to optimizing this protocol over the years. We thank Prof. Atsushi Miyawaki, RIKEN, Wako-city, Japan, for providing the FUCCI constructs. N.K.H. is a Cameron fellow of the Melanoma and Skin Cancer Research Institute, Australia. K.A.B. is a fellow of Cancer Institute New South Wales (13/ECF/1-39). The work leading to this protocol was supported by project grants RG 09-08 and RG 13-06 (Cancer Council New South Wales), 570778 and 1051996 (Priority-driven collaborative cancer research scheme/Cancer Australia/Cure Cancer Australia Foundation), 08/RFG/1-27 (Cancer Institute New South Wales), APP1003637 and APP1084893 (National Health and Medical Research Council).
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Spoerri, L., Beaumont, K.A., Anfosso, A., Haass, N.K. (2017). Real-Time Cell Cycle Imaging in a 3D Cell Culture Model of Melanoma. In: Koledova, Z. (eds) 3D Cell Culture. Methods in Molecular Biology, vol 1612. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7021-6_29
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DOI: https://doi.org/10.1007/978-1-4939-7021-6_29
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