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Methods Mol Biol. 2009;482:21-33. doi: 10.1007/978-1-59745-060-7_2.

Patterning mouse and human embryonic stem cells using micro-contact printing.

Author information

1
Terrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.

Abstract

Local micro-environmental cues consisting of soluble cytokines, extra-cellular matrix (ECM), and cell-cell contacts are determining factors in stem cell fate. These extrinsic cues form a 'niche' that governs a stem cell's decision to either self-renew or differentiate into one or more cell types. Recently, it has been shown that micro-patterning stem cells in two- and three-dimensions can provide direct control over several parameters of the local micro-environment, including colony size, distance between colonies, ECM substrate, and homotypic or heterotypic cell-cell contact. The protocol described here uses micro-contact printing to pattern ECM onto tissue culture substrates. Cells are seeded onto the patterned substrates in serum-free media and are confined to the patterned features. After patterning, stem cell phenotype is analyzed using quantitative immunocytochemistry and immunohistochemistry.

PMID:
19089347
DOI:
10.1007/978-1-59745-060-7_2
[Indexed for MEDLINE]

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