Abstract:
:Glass microfluidic devices have been fabricated to monitor the secretion of glycerol or fatty acids from cultured murine 3T3-L1 adipocytes. In the current studies, adipocytes are perfused in a reversibly sealed cell chamber, and secreted products are analyzed by enzyme assay on either a single- or dual-chip device. The analysis of glycerol employed the use of a dual-chip system, which used separate chips for cell perfusion and sample analysis. An improved single-chip device integrated the cell perfusion chamber and analysis component on one platform. The performance of this device was demonstrated by the analysis of fatty acids but could also be applied to analysis of glycerol or other chemicals. The single-chip system required fewer cells and lower flow rates and provided improved temporal response. In both systems, cells were perfused with buffer to monitor basal response followed by lipolysis stimulation with the β-adrenergic agonist isoproterenol. Measured basal glycerol concentration from 50,000 cells was 28 μM, and when stimulated, a spike threefold higher than basal concentration was detected followed by a continuous release 40% above basal levels. Fatty acid basal concentration was 24 μM, measured from 6200 cells, and isoproterenol stimulation resulted in a constant elevated concentration sevenfold higher than basal levels.
journal_name
Methods Enzymoljournal_title
Methods in enzymologyauthors
Dugan CE,Kennedy RTdoi
10.1016/B978-0-12-800280-3.00011-6subject
Has Abstractpub_date
2014-01-01 00:00:00pages
195-209eissn
0076-6879issn
1557-7988pii
B978-0-12-800280-3.00011-6journal_volume
538pub_type
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