Protocol

Stem Cells in Regenerative Medicine

Volume 482 of the series Methods in Molecular Biology pp 21-33

Patterning Mouse and Human Embryonic Stem Cells Using Micro-contact Printing

  • Raheem PeeraniAffiliated withTerrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of Toronto
  • , Celine BauwensAffiliated withTerrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of Toronto
  • , Eugenia KumachevaAffiliated withDepartment of Chemistry, University of Toronto
  • , Peter W. ZandstraAffiliated withInstitute of Biomaterials and Biomedical Engineering, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto

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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.

Key words

Micro-contact printing soft lithography embryonic stem cell flow cytometry quantitative immunohistochemistry ECM–cell interactions high throughput screening