Patterning Mouse and Human Embryonic Stem Cells Using Micro-contact Printing
Part of the
Methods in Molecular Biology
book series (MIMB, volume 482)
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 wordsMicro-contact printing soft lithography embryonic stem cell flow cytometry quantitative immunohistochemistry ECM–cell interactions high throughput screening
Flaim, C.J., S. Chien, and S.N. Bhatia, (2005) An extracellular matrix microarray for probing cellular differentiation.
Nat Methods, 2
(2): p. 119–25.CrossRefPubMedGoogle Scholar
McBeath, R., et al., (2004) Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment.
Dev Cell, 6
(4): p. 483–95.CrossRefPubMedGoogle Scholar
Soen, Y., et al., (2006) Exploring the regulation of human neural precursor cell differentiation using arrays of signaling microenvironments.
Mol Syst Biol, 2
: p. 37.CrossRefPubMedGoogle Scholar
Tan, J.L., et al., (2004) Simple approach to micropattern cells on common culture substrates by tuning substrate wettability.
Tissue Eng, 10
(5–6): p. 865–72.CrossRefPubMedGoogle Scholar
Chen, C.S., et al., (1997) Geometric control of cell life and death.
(5317): p. 1425–8.CrossRefPubMedGoogle Scholar
Anderson, D.G., S. Levenberg, and R. Langer, (2004) Nanoliter-scale synthesis of arrayed biomaterials and application to human embryonic stem cells.
Nat Biotechnol, 22
(7): p. 863–6.CrossRefPubMedGoogle Scholar
Rettig, J.R. and A. Folch, Large-scale single-cell trapping and imaging using microwell arrays.
Anal Chem, 2005. 77
(17): p. 5628–34.(2004) Molded polyethylene glycol microstructures for capturing cells within microfluidic channels.
Lab Chip, 4
(5): p. 425–30.Google Scholar
Folch, A., et al., (1999) Molding of deep polydimethylsiloxane microstructures for microfluidics and biological applications.
J Biomech Eng, 121
(1): p. 28–34.CrossRefPubMedGoogle Scholar
Huang, Y., et al., (2001) Electric manipulation of bioparticles and macromolecules on microfabricated electrodes.
Anal Chem, 73
(7): p. 1549–59.CrossRefPubMedGoogle Scholar
Kane, R.S., et al., (1999) Patterning proteins and cells using soft lithography.
(23–24): p. 2363–76.CrossRefPubMedGoogle Scholar
© Humana Press, a part of Springer Science+Business Media, LLC 2009