Abstract
The study of dynamic processes in the bone metastatic compartment has been challenged by the restrictive access and limited live imaging capabilities that in vivo bone models provide. In this protocol, we show the use of a human bone metastatic bioengineered microtissue for the quantitative investigation of cancer cells in an in vitro bone-like microenvironment. Using live cell epifluorescence microscopy, traditional- and spinning disc-confocal laser scanning microscopy, we demonstrate how to obtain multidimensional real-time data of fluorescently labeled cancer cells in the metastatic microenvironment. Using 4D imaging data processing software such as ImageJ and Imaris, we show how to transform qualitative images and videos into quantitative data of cancer cell attachment, morphology, proliferation, and migration in vitro in the human bone metastatic microtissue.
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Acknowledgements
This work was supported by an IHBI ECR grant, Advance Queensland (AQ) Maternity Fund Award from the Queensland Government (DSITI), Young Researcher Award (2017-YR-RoW-9) from Lush (UK), supporting non-animal testing alternatives, National Health and Medical Research Council (NHMRC) Peter Doherty Early Career Research Fellowship (RF) (APP1091734), and John Mills Young Investigator Award (YI0715) from the Prostate Cancer Foundation of Australia (PCFA). The author acknowledges the TRI for the excellent core facilities that enabled this research, and thank Adler Ju from the TRI microscopy facility. The author also thanks D/Prof. Dietmar W. Hutmacher, D/Prof. Judith A. Clements AC, Dr. Elena De-Juan-Pardo, Mr. Jose Malayil, Dr. Asha Mathew, Ms. Teja Yarlagadda, Dr. Sally Stephenson, Dr. Brett Hollier.
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Bock, N., Röhl, J. (2019). Real-Time and 3D Quantification of Cancer Cell Dynamics: Exploiting a Bioengineered Human Bone Metastatic Microtissue. In: Batra, J., Srinivasan, S. (eds) Theranostics. Methods in Molecular Biology, vol 2054. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9769-5_3
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DOI: https://doi.org/10.1007/978-1-4939-9769-5_3
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