Details:

Alternative Title:
Nat Protoc
Personal Author:
Fischer, Robert S ; Myers, Kenneth A ; Gardel, Margaret L ; Waterman, Clare M ;
Fischer, Robert S ; Myers, Kenneth A ; Gardel, Margaret L ; Waterman, Clare M ; Less -
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.
Subject:
[+]
Acrylic Resins
Article
Cell Communication
Cell Culture Techniques
Cellular Microenvironment
Extracellular Matrix
Human Umbilical Vein Endothelial Cells
Microscopy, Phase-Contrast

Source:
Nat Protoc. 2012 ; 7(11).;
Pubmed ID:
23099487
Pubmed Central ID:
PMC3845971
Document Type:
Journal Article ;
Funding:
DP10D00354/DP/NCCDPHP CDC HHS/United States ; Z99 HL999999/Intramural NIH HHS/United States ; ZIA HL005106-05/Intramural NIH HHS/United States ;
DP10D00354/DP/NCCDPHP CDC HHS/United States ; Z99 HL999999/Intramural NIH HHS/United States ; ZIA HL005106-05/Intramural NIH HHS/United States ; Less -
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:120a324f5543cc00ed0d33ea2d61c46ceac7f9ead3639130d81c4cd4abbd8baf
File Type:

Supporting Files

• 	NIHMS523235-supplement-Supplmental_Figure_1.pdf	pdf
  	nihms523235.nxml	txt
  	nihms523235f1.gif	gif
  	nihms523235f1.jpg	jpeg
  	nihms523235f2.gif	gif
  	nihms523235f2.jpg	jpeg

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