Document Detail


Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity.
MedLine Citation:
PMID:  21293460     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
We describe the use of a microfabricated cell culture substrate, consisting of a uniform array of closely spaced, vertical, elastomeric microposts, to study the effects of substrate rigidity on cell function. Elastomeric micropost substrates are micromolded from silicon masters comprised of microposts of different heights to yield substrates of different rigidities. The tips of the elastomeric microposts are functionalized with extracellular matrix through microcontact printing to promote cell adhesion. These substrates, therefore, present the same topographical cues to adherent cells while varying substrate rigidity only through manipulation of micropost height. This protocol describes how to fabricate the silicon micropost array masters (∼2 weeks to complete) and elastomeric substrates (3 d), as well as how to perform cell culture experiments (1-14 d), immunofluorescence imaging (2 d), traction force analysis (2 d) and stem cell differentiation assays (1 d) on these substrates in order to examine the effect of substrate rigidity on stem cell morphology, traction force generation, focal adhesion organization and differentiation.
Authors:
Michael T Yang; Jianping Fu; Yang-Kao Wang; Ravi A Desai; Christopher S Chen
Publication Detail:
Type:  Journal Article     Date:  2011-01-27
Journal Detail:
Title:  Nature protocols     Volume:  6     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2011-02-04     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  187-213     Citation Subset:  IM    
Affiliation:
1] Department of Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA. [2].
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