Abstract
Cells are surrounded by mechanical stimuli in their microenvironment. It is important to determine how cells respond to the mechanical information that surrounds them in order to understand both development and disease progression, as well as to be able to predict cell behavior in response to physical stimuli. Here we describe a protocol to determine the effects of interstitial fluid flow on the migratory behavior of an aggregate of epithelial cells in a three-dimensional (3D) culture model. This protocol includes detailed methods for the fabrication of a 3D cell culture chamber with hydrostatic pressure control, the culture of epithelial cells as an aggregate in a collagen gel, and the analysis of collective cell behavior in response to pressure-driven flow.
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Acknowledgements
This work was supported in part by grants from the NIH (GM083997, HL110335, and HL118532), pilot project funding from the NIH/NCI Physical Sciences-Oncology Center at Princeton University (U54CA143803), Concept Award W81XWH-09-1-0565 from the Breast Cancer Research Program of the Department of Defense (to J.T.), the David & Lucile Packard Foundation, the Alfred P. Sloan Foundation, the Camille & Henry Dreyfus Foundation, and the Burroughs Wellcome Fund.
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Piotrowski-Daspit, A.S., Simi, A.K., Pang, MF., Tien, J., Nelson, C.M. (2017). A 3D Culture Model to Study How Fluid Pressure and Flow Affect the Behavior of Aggregates of Epithelial Cells. In: Martin, F., Stein, T., Howlin, J. (eds) Mammary Gland Development. Methods in Molecular Biology, vol 1501. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6475-8_12
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DOI: https://doi.org/10.1007/978-1-4939-6475-8_12
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