title: A microfluidic cell culture system for monitoring of sequential changes in endothelial cells after heat stress.
abstract: Endothelial damage induced by a highly elevated body temperature is crucial in some diseases including viral hemorrhagic fevers. Here, we report the heat-induced sequential changes of endothelial cells under shear stress, which were determined with a microfluidic culture system. Although live cell imaging showed only minor changes in the appearance of heat-treated cells, Hsp70 mRNA expression analysis demonstrated that the endothelial cells in channels of the system responded well to heat treatment. F-actin staining also revealed clear changes in the bundles of actin filaments after heat treatment. Well-organized bundles of actin filaments in control cells disappeared in heat-treated cells cultured in the channel. Furthermore, the system enabled detection of sequential changes in plasminogen activator inhibitor-1 (PAI-1) secretion from endothelial cells. PAI-1 concentration in the effluent solution was significantly elevated for the first 15min after initiation of heat treatment, and then decreased subsequently. This study provides fundamental information on heat-induced endothelial changes under shear stress and introduces a potent tool for analyzing endothelial secretions.
authors.affiliation: Institute of Microchemical Technology Co. Ltd., 207 East KSP, 3-2-1 Sakado, Takatsu, Kawasaki, Kanagawa 213-0012, Japan.
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Tazawa
authors.affiliation: Department of Life Sciences, Toyo University, 1-1-1 Itakura, Oura, Gunma 374-0193, Japan.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Sato
authors.affiliation: Department of Life Sciences, Toyo University, 1-1-1 Itakura, Oura, Gunma 374-0193, Japan.
authors.forename: Atsuhiro
authors.initials: A
authors.lastname: Tsutiya
authors.affiliation: Division of Biotechnology and Macromolecular Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan; FIRST Research Center for Innovative Nanobiodevices and Innovative Research Center for Preventive Medical Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Tokeshi
authors.affiliation: Department of Life Sciences, Toyo University, 1-1-1 Itakura, Oura, Gunma 374-0193, Japan; Bio-Nano Electronic Research Centre, Toyo University, 2100 Kujirai, Kawagoe, Saitama 350-8585, Japan. Electronic address: r-kaneko@toyo.jp.
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Ohtani-Kaneko
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000222:Adaptation, Physiological / Q000502:physiology; D000818:Animals; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation*; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D000882:Haplorhini; D018869:Heat-Shock Response / Q000502:physiology*; D006361:Heating / Q000295:instrumentation*; D006358:Hot Temperature; D006801:Humans; D056656:Lab-On-A-Chip Devices; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism
chemical_list: D018840:HSP70 Heat-Shock Proteins; D017395:Plasminogen Activator Inhibitor 1; C051321:SERPINE1 protein, human
keywords: Endothelial cells; Heat-shock response; Microchip; Microfluidic cell culture system; PAI-1