Analysis of cyclic-stretching responses using cell-adhesion-patterned cells

Yuki Katanosaka, Jin Hua Bao, Tomoyo Komatsu, Tomohiko Suemori, Akira Yamada, Satoshi Mohri, Keiji Naruse

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)

Abstract

Human vascular endothelial cells form the interface between the bloodstream and vessel walls and are continuously subjected to mechanical stimulation. When endothelial cells are stretched cyclically, along one axis, they align perpendicular to the axis of stretch. We previously reported that applying a cyclic, uni-axial strain to cells induced tyrosine phosphorylation of focal adhesion kinase and stimulated mitogen-activated protein kinase. However, it is difficult to quantify and analyze the spatial distribution of tyrosine phosphorylation in these cells, as they form focal adhesions randomly. In this study, we developed a system to overcome this problem by preparing individual, uniform, patterned cells that could be stretched cyclically and uni-axially. We constructed polydimethylsiloxane stretch chambers and used microcontact printing technology to imprint a pattern of 2 μm fibronectin dots (10 lines × 10 columns in a 38 μm square) before seeding them with human umbilical vein endothelial cells (HUVEC). We found that most HUVEC attached to the patterned dots after 2 h and were similar in size and morphology, based on phase-contrast microscopy. In this system we were able to statistically analyze tyrosine phosphorylation and actin polymerization in these patterned cells, when subjected to a cyclic, uni-axial strain, using fluorescent microscopy.

Original languageEnglish
Pages (from-to)82-89
Number of pages8
JournalJournal of biotechnology
Volume133
Issue number1
DOIs
Publication statusPublished - Jan 1 2008

Keywords

  • HUVEC
  • Microcontact printing
  • PDMS stretch chamber
  • Tyrosine phosphorylation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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