title: Microfluidic perfusion culture chip providing different strengths of shear stress for analysis of vascular endothelial function.
abstract: We developed a microfluidic perfusion cell culture chip that provides three different shear stress strengths and a large cell culture area for the analysis of vascular endothelial functions. The microfluidic network was composed of shallow flow-control channels of three different depths and deep cell culture channels. The flow-control channels with high fluidic resistances created shear stress strengths ranging from 1.0 to 10.0 dyn/cm(2) in the cell culture channels. The large surface area of the culture channels enabled cultivation of a large number (approximately 6.0 × 10(5)) of cells. We cultured human umbilical vein endothelial cells (HUVECs) and evaluated the changes in cellular morphology and gene expression in response to applied shear stress. The HUVECs were aligned in the direction of flow when exposed to a shear stress of 10.0 dyn/cm(2). Compared with conditions of no shear stress, endothelial nitric oxide synthase mRNA expression increased by 50% and thrombomodulin mRNA expression increased by 8-fold under a shear stress of 9.5 dyn/cm(2).
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Hattori
authors.affiliation: Frontier Research Laboratories, R&D Division, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Munehira
authors.affiliation: Frontier Research Laboratories, R&D Division, Daiichi Sankyo Co., Ltd., 1-2-58, Hiromachi, Shinagawa-ku, Tokyo 140-8710, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Satoh
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan. Electronic address: shinji.sugiura@aist.go.jp.
authors.forename: Shinji
authors.initials: S
authors.lastname: Sugiura
authors.affiliation: Research Center for Stem Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan.
authors.forename: Toshiyuki
authors.initials: T
authors.lastname: Kanamori
mesh_terms: D002452:Cell Count; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000235:genetics*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D052250:Nitric Oxide Synthase Type III / Q000235:genetics / Q000378:metabolism; D010477:Perfusion; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D033081:Shear Strength; D013314:Stress, Mechanical; D018180:Thrombomodulin / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; C546931:THBD protein, human; D018180:Thrombomodulin; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III
keywords: Endothelial cell; Microfluidic device; Perfusion culture; Quantitative PCR; Shear stress