A multiwell platform for studying stiffness-dependent cell biology

Justin D. Mih, Asma S. Sharif, Fei Liu, Aleksandar Marinkovic, Matthew M. Symer, Daniel J Tschumperlin

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

Adherent cells are typically cultured on rigid substrates that are orders of magnitude stiffer than their tissue of origin. Here, we describe a method to rapidly fabricate 96 and 384 well platforms for routine screening of cells in tissue-relevant stiffness contexts. Briefly, polyacrylamide (PA) hydrogels are cast in glass-bottom plates, functionalized with collagen, and sterilized for cell culture. The Young's modulus of each substrate can be specified from 0.3 to 55 kPa, with collagen surface density held constant over the stiffness range. Using automated fluorescence microscopy, we captured the morphological variations of 7 cell types cultured across a physiological range of stiffness within a 384 well plate. We performed assays of cell number, proliferation, and apoptosis in 96 wells and resolved distinct profiles of cell growth as a function of stiffness among primary and immortalized cell lines. We found that the stiffness-dependent growth of normal human lung fibroblasts is largely invariant with collagen density, and that differences in their accumulation are amplified by increasing serum concentration. Further, we performed a screen of 18 bioactive small molecules and identified compounds with enhanced or reduced effects on soft versus rigid substrates, including blebbistatin, which abolished the suppression of lung fibroblast growth at 1 kPa. The ability to deploy PA gels in multiwell plates for high throughput analysis of cells in tissue-relevant environments opens new opportunities for the discovery of cellular responses that operate in specific stiffness regimes.

Original languageEnglish (US)
Article numbere19929
JournalPLoS One
Volume6
Issue number5
DOIs
StatePublished - 2011
Externally publishedYes

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Cytology
cell biology
Cell Biology
Collagen
Stiffness
Cultured Cells
collagen
Growth
Fibroblasts
polyacrylamide
Lung
Elastic Modulus
fibroblasts
cells
Tissue
Fluorescence Microscopy
lungs
Glass
Substrates
Cell Culture Techniques

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

A multiwell platform for studying stiffness-dependent cell biology. / Mih, Justin D.; Sharif, Asma S.; Liu, Fei; Marinkovic, Aleksandar; Symer, Matthew M.; Tschumperlin, Daniel J.

In: PLoS One, Vol. 6, No. 5, e19929, 2011.

Research output: Contribution to journalArticle

Mih, Justin D. ; Sharif, Asma S. ; Liu, Fei ; Marinkovic, Aleksandar ; Symer, Matthew M. ; Tschumperlin, Daniel J. / A multiwell platform for studying stiffness-dependent cell biology. In: PLoS One. 2011 ; Vol. 6, No. 5.
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