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J Vis Exp. 2016 May 9;(111). doi: 10.3791/53989.

Preparation of 3D Collagen Gels and Microchannels for the Study of 3D Interactions In Vivo.

Author information

1
Department of Cell and Regenerative Biology, University of Wisconsin-Madison; Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison; bmburkel@wisc.edu.
2
Department of Cell and Regenerative Biology, University of Wisconsin-Madison.
3
Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison; Department of Biomedical Engineering, University of Wisconsin-Madison; Morgridge Institute for Research, University of Wisconsin-Madison.
4
Department of Cell and Regenerative Biology, University of Wisconsin-Madison; Laboratory for Optical and Computational Instrumentation, University of Wisconsin-Madison; Paul P. Carbone Comprehensive Cancer center, University of Wisconsin-Madison.

Abstract

Historically, most cellular processes have been studied in only 2 dimensions. While these studies have been informative about general cell signaling mechanisms, they neglect important cellular cues received from the structural and mechanical properties of the local microenvironment and extracellular matrix (ECM). To understand how cells interact within a physiological ECM, it is important to study them in the context of 3 dimensional assays. Cell migration, cell differentiation, and cell proliferation are only a few processes that have been shown to be impacted by local changes in the mechanical properties of a 3-dimensional ECM. Collagen I, a core fibrillar component of the ECM, is more than a simple structural element of a tissue. Under normal conditions, mechanical cues from the collagen network direct morphogenesis and maintain cellular structures. In diseased microenvironments, such as the tumor microenvironment, the collagen network is often dramatically remodeled, demonstrating altered composition, enhanced deposition and altered fiber organization. In breast cancer, the degree of fiber alignment is important, as an increase in aligned fibers perpendicular to the tumor boundary has been correlated to poorer patient prognosis(1). Aligned collagen matrices result in increased dissemination of tumor cells via persistent migration(2,3). The following is a simple protocol for embedding cells within a 3-dimensional, fibrillar collagen hydrogel. This protocol is readily adaptable to many platforms, and can reproducibly generate both aligned and random collagen matrices for investigation of cell migration, cell division, and other cellular processes in a tunable, 3-dimensional, physiological microenvironment.

PMID:
27213771
PMCID:
PMC4942088
DOI:
10.3791/53989
[Indexed for MEDLINE]
Free PMC Article

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