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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Lineage*
  • Colony-Forming Units Assay
  • Dimethylpolysiloxanes
  • Embryonic Stem Cells / cytology*
  • Extracellular Matrix / metabolism
  • Humans
  • Immunohistochemistry
  • Mice
  • Solutions

Substances

  • Dimethylpolysiloxanes
  • Solutions