Microcontact Peeling: A Cell Micropatterning Technique for Circumventing Direct Adsorption of Proteins to Hydrophobic PDMS

Sho Yokoyama1, Tsubasa S. Matsui2, Shinji Deguchi2

1 Current: Micro/Nano Technology Center, Tokai University, Hiratsuka, 2 Current: Division of Bioengineering, Graduate School of Engineering Science, Osaka University, Osaka
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 10.21
DOI:  10.1002/cpcb.22
Online Posting Date:  June, 2017
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Abstract

Microcontact printing (μCPr) is one of the most popular techniques used for cell micropatterning. In conventional μCPr, a polydimethylsiloxane (PDMS) stamp with microfeatures is used to adsorb extracellular matrix (ECM) proteins onto the featured surface and transfer them onto particular areas of a cell culture substrate. However, some types of functional proteins other than ECM have been reported to denature upon direct adsorption to hydrophobic PDMS. Here we describe a detailed protocol of an alternative technique––microcontact peeling (μCPe)––that allows for cell micropatterning while circumventing the step of adsorbing proteins to bare PDMS. This technique employs microfeatured materials with a relatively high surface energy such as copper, instead of using a microfeatured PDMS stamp, to peel off a cell‐adhesive layer present on the surface of substrates. Consequently, cell‐nonadhesive substrates are exposed at the specific surface that undergoes the physical contact with the microfeatured material. Thus, although μCPe and μCPr are apparently similar, the former does not comprise a process of transferring biomolecules through hydrophobic PDMS. © 2017 by John Wiley & Sons, Inc.

Keywords: cell micropatterning; protein micropatterning; microcontact printing; microcontact peeling

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Microcontact Peeling (μCPe)
  • Support Protocol 1: Cell Culture
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Microcontact Peeling (μCPe)

  Materials
  • Sylgard 184 Silicone Elastomer Kit (e.g., Dow Corning) containing:
    • Base polymer
    • Cross linker
  • Pluronic F‐127 (e.g., Invitrogen)
  • Phosphate‐buffered saline (PBS; see appendix 2A)
  • Gelatin (e.g., Sigma‐Aldrich)
  • Cells of interest (see protocol 2Support Protocol)
  • Vacuum chamber
  • 35‐mm glass‐bottom or polystyrene cell culture dish
  • Spin coater (e.g., Kyowa Riken K‐359S1)
  • 65°C oven
  • Glow discharge plasma generator (e.g., Meiwafosis SEDE‐P or SEDE‐GE)
  • Copper electron microscopy grids (e.g., EM Japan, cat. no. G203)
  • Vacuum tweezer (e.g., Virtual Industries Tweezer‐Vac)
  • Fine tweezers
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Figures

Videos

Literature Cited

Literature Cited
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