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

  • Raheem Peerani
  • Celine Bauwens
  • Eugenia Kumacheva
  • Peter W. Zandstra
Part of the Methods in Molecular Biology book series (MIMB, volume 482)

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 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Raheem Peerani
    • 1
  • Celine Bauwens
    • 2
  • Eugenia Kumacheva
    • 3
  • Peter W. Zandstra
    • 4
  1. 1.Terrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of TorontoTorontoCanada
  2. 2.Terrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of TorontoTorontoCanada
  3. 3.Department of ChemistryUniversity of TorontoTorontoCanada
  4. 4.Institute of Biomaterials and Biomedical Engineering, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of TorontoTorontoCanada

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