Microbioassay system for antiallergic drug screening using suspension cells retaining in a poly(dimethylsiloxane) microfluidic device.
This article describes an antiallergic drug-screening system by the detection of histamine released from mast cells (suspension cells) on a multilayer microchip. In this study, the elastmeric material, poly(dimethylsiloxane) (PDMS), was employed to fabricate microchannels and microchambers. The microchip consists of two sections: a histamine-releasing one, which has a cell chamber, and a histamine-derivatizing one. Both were laminated to one microchip. Rat peritoneal mast cells were retained in the cell chamber (1.2 microL) with a filtering system using a cellulose nitrate membrane. This filtering system could easily retain suspension cells without cell damage. Mast cells were viable for a sufficient time to conduct the assay on the cell chamber. The cells were stimulated with a chemical release compound 48/80 (C48/80), and then histamine flowed into the lower layer, where it was derivatized to the fluorescent molecules with o-phthalaldehyde and its fluorescence was detected on the microchip. This flow system could detect the time course of the histamine release, and this microchip system required only 20 min for the assay. By this integrated system, 51 pmol of histamine released from 500 cells was detected, and the number of cells required for the assay was reduced to 1% compared with conventional bulk systems. By comparing the released histamine levels with and without drugs, their effect could be evaluated. The inhibition ratio of C48/80 induced-histamine release using an antiallergic drug, disodium cromoglicate (DSCG), was related to the concentration of DSCG. This flow system was applicable for antiallergy drug screening by rapid measurement of the inhibition of histamine release from a very small amount of mast cells.
Tokuyama
Takahito
T
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi, Bunkyo, Tokyo 113-8657, Japan.
Fujii
Shin-Ichiro
S
Sato
Kiichi
K
Abo
Mitsuru
M
Okubo
Akira
A
D000818:Animals; D018926:Anti-Allergic Agents / Q000032:analysis* / Q000494:pharmacology; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D002478:Cells, Cultured; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D005453:Fluorescence; D006632:Histamine / Q000032:analysis*; D006636:Histamine Release; D008407:Mast Cells / Q000378:metabolism; D008832:Microchemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D013997:Time Factors; D009764:o-Phthalaldehyde / Q000737:chemistry
D018926:Anti-Allergic Agents; D004129:Dimethylpolysiloxanes; D009764:o-Phthalaldehyde; D006632:Histamine