Authors: Anna M Clark Kyle M Sousa Claire N Chisolm Ormond A MacDougald Robert T Kennedy
Publish Date: 2010/06/12
Volume: 397, Issue: 7, Pages: 2939-2947
Abstract
A threelayer microfluidic device was developed that combined perfusion of cultured cells with online chemical analysis for near realtime monitoring of cellular secretions Two layers were reversibly sealed to form a cell chamber that allowed cells grown on coverslips to be loaded directly into the chip The outlet of the chamber was in fluidic contact with a third layer that was permanently bonded Perfusate from the cell chamber flowed into this third layer where a fluorescence enzyme assay for nonesterified fatty acid NEFA was performed online The device was used to monitor efflux of NEFAs from ∼6200 cultured adipocytes with 83 s temporal resolution Perfusion of murine 3T3L1 cultured adipocytes resulted in an average basal concentration of 242 ± 24 μM NEFA SEM n = 6 detected in the effluent corresponding to 331 × 10−5 nmol cell−1 min−1 Upon pharmacological treatment with a βadrenergic agonist to stimulate lipolysis a 69 ± 07fold SEM n = 6 sustained increase in NEFA secretion was observed This multilayer device provides a versatile platform that could be adapted for use with other cell types to study corresponding cellular secretions in near realtime
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