title: Spatial Chemical Stimulation Control in Microenvironment by Microfluidic Probe Integrated Device for Cell-Based Assay.
abstract: Cell-cell interactions play an important role in the development and function of multicellular organisms. To investigate these interactions in detail, it is necessary to evaluate the behavior of a cell population when the minimum number of cells in the population is stimulated by some chemical factors. We propose a microfluidic device integrated with microfluidic probe (MFP) functionality; this device is capable of imparting a chemical stimulus to cells within a microenvironment, for cell-based assays. The device contains MFP channels at the walls of the cell culture microchannels, and it can control a localized chemical stimulation area at the scale of a single cell to a few cells using MFP fluid control in a microspace. The results of a finite element method-based simulation indicated that it is possible to control the chemical stimulation area at the scale of a single cell to a few cells by optimizing the MFP channel apex width and the flow ratio. In addition, localized cell staining was demonstrated successfully using a spatial chemical stimulus. We confirmed the device functionality as a novel cell-based assay tool. We succeeded in performing localized cell collection using this method, which suggested that the single cell analysis of a cell monolayer that is subjected to a specific chemical stimulus is possible. The method proposed in this paper can contribute significantly to the fields of cell biology and drug development.
authors.affiliation: Department of Mechanical Engineering, Tokai University, Hiratsuka, Kanagawa, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Horayama
authors.affiliation: Department of Mechanical Engineering, Tokai University, Hiratsuka, Kanagawa, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Shinha
authors.affiliation: Department of Chemistry, Osaka University, Toyonaka, Osaka, Japan.|Micro/Nano Technology Center, Tokai University, Hiratsuka, Kanagawa, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Kabayama
authors.affiliation: Institute of Industrial Science, The University of Tokyo, Meguro, Tokyo, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Fujii
authors.affiliation: Department of Mechanical Engineering, Tokai University, Hiratsuka, Kanagawa, Japan.|Micro/Nano Technology Center, Tokai University, Hiratsuka, Kanagawa, Japan.
authors.forename: Hiroshi
authors.identifier: http://orcid.org/0000-0003-0623-0248
authors.initials: H
authors.lastname: Kimura
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000737:chemistry / Q000187:drug effects; D002478:Cells, Cultured / Q000737:chemistry* / Q000187:drug effects; D060833:Cellular Microenvironment; D017076:Computer-Aided Design; D003412:Cricetulus; D020342:Finite Element Analysis; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D012235:Rhodamines / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
chemical_list: D012235:Rhodamines; D014357:Trypsin; C029773:rhodamine B