Stiffness-controlled three-dimensional extracellular matrices for high-resolution imaging of cell behavior

Fischer, Robert S; Myers, Kenneth A; Gardel, Margaret L; Waterman, Clare M

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Stiffness-controlled three-dimensional extracellular matrices for high-resolution imaging of cell behavior

Published Date:

Oct 25 2012

Source:

Nat Protoc. 2012; 7(11).

[PDF-1.61 MB]

Details:

Personal Authors:

Fischer, Robert S ; Myers, Kenneth A ; Gardel, Margaret L ; Waterman, Clare M ;

Keywords:

[+]

Pubmed ID:

23099487

Pubmed Central ID:

PMC3845971

Description:

Regulation of cell functions by the physical properties of the extracellular matrix (ECM) has emerged as a crucial contributor to development and disease. Two specific physical properties of the ECM, stiffness and dimensionality, each influence cell signaling and function. As these ECM physical properties are linked to other properties that also regulate cell behavior, e.g., integrin ligand density, parsing the specific contributions of ECM stiffness and dimensionality has proven difficult. Here we detail a simple protocol, which can be completed in 1-2 d, for combining three-dimensional (3D) ECM engagement with controlled underlying ECM stiffness. In these 'sandwich gels', cells are sandwiched between a 3D fibrillar ECM and an ECM-coupled polyacrylamide gel of defined compliance, allowing the study of the specific effects of ECM compliance on cell function in physiologically relevant 3D ECMs. This type of system enables high-resolution time-lapse imaging and is suitable for a wide range of cell types and molecular perturbations.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

DP10D00354/DP/NCCDPHP CDC HHS/United States
Z99 HL999999/Intramural NIH HHS/United States
ZIA HL005106-05/Intramural NIH HHS/United States

Main Document Checksum:

[+]

Supporting Files:

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	nihms523235.nxml	text/plain
	nihms523235f1.gif	image/gif
	nihms523235f1.jpg	image/jpeg
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