Patterning mouse and human embryonic stem cells using micro-contact printing.
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.
Peerani
Raheem
R
Terrence Donnelly Centre for Cellular and Biomolecular Research, Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada.
Bauwens
Celine
C
Kumacheva
Eugenia
E
Zandstra
Peter W
PW
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D019070:Cell Lineage; D003114:Colony-Forming Units Assay; D004129:Dimethylpolysiloxanes; D053595:Embryonic Stem Cells / Q000166:cytology*; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D012996:Solutions
D004129:Dimethylpolysiloxanes; D012996:Solutions