Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity

Michael T. Yang, Jianping Fu, Yang Kao Wang, Ravi A. Desai, Christopher S. Chen

研究成果: Article

167 引文 (Scopus)

摘要

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.

原文English
頁(從 - 到)187-213
頁數27
期刊Nature Protocols
6
發行號2
DOIs
出版狀態Published - 2011 二月 1

指紋

Biophysics
Stem cells
Rigidity
Stem Cells
Traction
Silicon
Substrates
Cell Culture Techniques
Colony-Forming Units Assay
Printing
Focal Adhesions
Cell Adhesion
Cues
Fluorescent Antibody Technique
Extracellular Matrix
Cell culture
Cell Differentiation
elastomeric
Traction (friction)
Cell adhesion

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

引用此文

Yang, Michael T. ; Fu, Jianping ; Wang, Yang Kao ; Desai, Ravi A. ; Chen, Christopher S. / Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity. 於: Nature Protocols. 2011 ; 卷 6, 編號 2. 頁 187-213.
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Assaying stem cell mechanobiology on microfabricated elastomeric substrates with geometrically modulated rigidity. / Yang, Michael T.; Fu, Jianping; Wang, Yang Kao; Desai, Ravi A.; Chen, Christopher S.

於: Nature Protocols, 卷 6, 編號 2, 01.02.2011, p. 187-213.

研究成果: Article

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