title: A pneumatic pressure-driven multi-throughput microfluidic circulation culture system.
abstract: Here, we report a pneumatic pressure-driven microfluidic device capable of multi-throughput medium circulation culture. The circulation culture system has the following advantages for application in drug discovery: (i) simultaneous operation of multiple circulation units, (ii) use of a small amount of circulating medium (3.5 mL), (iii) pipette-friendly liquid handling, and (iv) a detachable interface with pneumatic pressure lines via sterile air-vent filters. The microfluidic device contains three independent circulation culture units, in which human umbilical vein endothelial cells (HUVECs) were cultured under physiological shear stress induced by circulation of the medium. Circulation of the medium in the three culture units was generated by programmed sequentially applied pressure from two pressure-control lines. HUVECs cultured in the microfluidic device were aligned under a one-way circulating flow with a shear stress of 10 dyn cm(-2); they exhibited a randomly ordered alignment under no shear stress and under reciprocating flow with a shear stress of 10 dyn cm(-2). We also observed 2.8- to 4.9-fold increases in expression of the mRNAs of endothelial nitric oxide synthase and thrombomodulin under one-way circulating flow with a shear stress of 10 dyn cm(-2) compared with conditions of no shear stress or reciprocating flow.
authors.affiliation: Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Central 5th, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan. shinji.sugiura@aist.go.jp.
authors.forename: T
authors.initials: T
authors.lastname: Satoh
authors.forename: G
authors.initials: G
authors.lastname: Narazaki
authors.forename: R
authors.initials: R
authors.lastname: Sugita
authors.forename: H
authors.initials: H
authors.lastname: Kobayashi
authors.forename: S
authors.initials: S
authors.lastname: Sugiura
authors.forename: T
authors.initials: T
authors.lastname: Kanamori
mesh_terms: D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D003470:Culture Media; D005452:Fluoresceins; D005456:Fluorescent Dyes; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation*; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D013314:Stress, Mechanical; D018180:Thrombomodulin / Q000235:genetics
chemical_list: D003470:Culture Media; D005452:Fluoresceins; D005456:Fluorescent Dyes; D018180:Thrombomodulin; C085925:calcein AM; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III