Microfluidic genipin deposition technique for extended culture of micropatterned vascular muscular thin films

Eric S. Hald, Kerianne E. Steucke, Jack A. Reeves, Zaw Win, Patrick W. Alford

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

The chronic nature of vascular disease progression requires the development of experimental techniques that simulate physiologic and pathologic vascular behaviors on disease-relevant time scales. Previously, microcontact printing has been used to fabricate two-dimensional functional arterial mimics through patterning of extracellular matrix protein as guidance cues for tissue organization. Vascular muscular thin films utilized these mimics to assess functional contractility. However, the microcontact printing fabrication technique used typically incorporates hydrophobic PDMS substrates. As the tissue turns over the underlying extracellular matrix, new proteins must undergo a conformational change or denaturing in order to expose hydrophobic amino acid residues to the hydrophobic PDMS surfaces for attachment, resulting in altered matrix protein bioactivity, delamination, and death of the tissues. Here, we present a microfluidic deposition technique for patterning of the crosslinker compound genipin. Genipin serves as an intermediary between patterned tissues and PDMS substrates, allowing cells to deposit newly-synthesized extracellular matrix protein onto a more hydrophilic surface and remain attached to the PDMS substrates. We also show that extracellular matrix proteins can be patterned directly onto deposited genipin, allowing dictation of engineered tissue structure. Tissues fabricated with this technique show high fidelity in both structural alignment and contractile function of vascular smooth muscle tissue in a vascular muscular thin film model. This technique can be extended using other cell types and provides the framework for future study of chronic tissue- and organ-level functionality.

Original languageEnglish (US)
Article numbere52971
JournalJournal of Visualized Experiments
Volume2015
Issue number100
DOIs
StatePublished - Jun 26 2015

Keywords

  • Arterial tissue engineering
  • Bioengineering
  • Cell viability
  • Genipin
  • In vitro disease model
  • Issue 100
  • Mechanical properties
  • Microfluidics
  • Polydimethylsiloxane
  • Protein deposition
  • Substrate modification
  • Vascular smooth muscle cells

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