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Wensaas 2007 J Lipid Res

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Publications in the MiPMap
Wensaas AJ, Rustan AC, Lövstedt K, Kull B, Wikström S, Drevon CA, Hallén S (2007) Cell-based multiwell assays for the detection of substrate accumulation and oxidation. J Lipid Res 48:961-7.

» PMID: 17213484 Open Access

Wensaas AJ, Rustan AC, Loevstedt K, Kull B, Wikstroem S, Drevon CA, Hallen S (2007) J Lipid Res

Abstract: We describe multiwell assays for detecting the accumulation as well as the subsequent oxidation of (14)C-labeled substrates in cultured cells. Accumulation is monitored in real time by an established scintillation proximity assay in which the scintillator is embedded in the plate base primarily detecting cell-associated radiolabel. The substrate oxidation assay is a novel variant of previously described experimental approaches aimed at trapping (14)CO(2) produced by isolated enzymes, organelles, or intact cells. This method uses a standard 96-well tissue culture plate and, on top, an inverted filter plate immersed with NaOH that are clamped into a sandwich sealed with a silicon gasket to obtain gas-tight compartments. (14)CO(2) is captured in the filter and quantified by conventional scintillation. We demonstrate both the accumulation and subsequent oxidation of (14)C-labeled substrates in cultured human myotubes, adipocytes, and hepatocytes. Both methods are adaptable for compound screening; at the same time, these protocols provide easy-to-use and time- saving methods for in vitro studies of cellular fuel handling. Keywords: Fatty acids, Glucose uptake, CO2 capture, Scintillation proximity assay

O2k-Network Lab: SE Molndal Hallen S


Organism: Human  Tissue;cell: Skeletal muscle, Liver, Fat