title: Analysis of cyclic-stretching responses using cell-adhesion-patterned cells.
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 microm fibronectin dots (10 lines x 10 columns in a 38 microm square) before seeding them with human umbilical vein endothelial cells (HUVEC). We found that most HUVEC attached to the patterned dots after 2h 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.
authors.affiliation: Department of Cardiovascular Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Katanosaka
authors.forename: Jin-Hua
authors.initials: JH
authors.lastname: Bao
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Komatsu
authors.forename: Tomohiko
authors.initials: T
authors.lastname: Suemori
authors.forename: Akira
authors.initials: A
authors.lastname: Yamada
authors.forename: Satoshi
authors.initials: S
authors.lastname: Mohri
authors.forename: Keiji
authors.initials: K
authors.lastname: Naruse
mesh_terms: D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D016764:Cell Polarity / Q000187:drug effects; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000187:drug effects / Q000502:physiology*; D009757:Nylons / Q000737:chemistry*
chemical_list: D004129:Dimethylpolysiloxanes; D005353:Fibronectins; D009757:Nylons; C085707:poly(dimethylsiloxane)-polyamide copolymer