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

Methods in molecular biology (Clifton, N.J.) (2008-12-18)
Raheem Peerani, Celine Bauwens, Eugenia Kumacheva, Peter W Zandstra
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Mouse IgG (Fc specific) F(ab′)2 fragment−FITC antibody produced in goat, flow cytometry grade, affinity isolated antibody, buffered aqueous solution

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