Reversibly sealed multilayer microfluidic device for integrated cell perfusion and on-line chemical analysis of cultured adipocyte secretions |
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Authors: | Anna M Clark Kyle M Sousa Claire N Chisolm Ormond A MacDougald Robert T Kennedy |
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Institution: | (1) Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA;(2) Department of Molecular & Integrative Physiology, University of Michigan, Ann Arbor, MI 48109, USA;(3) Department of Internal Medicine, University of Michigan, Ann Arbor, MI 48109, USA;(4) Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA; |
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Abstract: | A three-layer microfluidic device was developed that combined perfusion of cultured cells with on-line chemical analysis for
near real-time 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 non-esterified fatty acid (NEFA) was performed on-line. The device was used to monitor efflux of NEFAs from ∼6,200 cultured
adipocytes with 83 s temporal resolution. Perfusion of murine 3T3-L1 cultured adipocytes resulted in an average basal concentration
of 24.2 ± 2.4 μM NEFA (SEM, n = 6) detected in the effluent corresponding to 3.31 × 10−5 nmol cell−1 min−1. Upon pharmacological treatment with a β-adrenergic agonist to stimulate lipolysis, a 6.9 ± 0.7-fold (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 real-time. |
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