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A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells

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dc.creatorDiaz, Paola A. Riveraspa
dc.creatorCamargo, Doris E. Gómezspa
dc.creatorOndo Méndez, Alejandro Oyonospa
dc.creatorGómez-Alegría, Claudio J.spa
dc.date.accessioned2020-05-26T00:09:56Z
dc.date.available2020-05-26T00:09:56Z
dc.date.created2020spa
dc.description.abstractIntroduction: The quantitation of glucose consumption in animal cell cultures is mainly based on the use of radiolabeled or fluorescent analogues, resulting in expensive and tedious procedures, requiring special equipment and, sometimes, with potential health and environmental risks. Objectives: The objective of this work was to evaluate the application of a blood plasma colorimetric assay to quantify glucose consumption in in vitro cultures of adipose cells. Methods: We worked with 3T3-L1 adipose cells differentiated by 7–8 days, which were exposed to different initial glucose concentrations (5.5, 2.8 and 1.4 mM) for variable times, either in the absence or the presence of 100 nM insulin. Using a commercial colorimetric glucose assay, extracellular glucose was determined, and glucose uptake was calculated as the difference between the initial and final glucose concentration. Results: The colorimetric assay allowed us to quantify glucose uptake in our cell model, observing a linear response over time (r2?0.9303) to the different glucose concentrations, both in the basal and insulin-induced condition. The insulin-stimulated glucose consumption was higher than basal consumption at all glucose concentrations evaluated, but significant differences were observed at 120-, 360- and 480-min in glucose 5.5 mM (p ? 0.01, n = 5), and 240 min in glucose 1.4 mM (p ? 0.01, n = 5). A Vmax of 4.1 and 5.9 nmol/ml/min (basal and insulin-induced, respectively) and a Km of 1.1 mM (same in basal vs insulin-stimulated) were calculated. The bioassay was also useful in a pharmacological context: in glucose 1.4 mM, glucose consumption showed an effect that depended on insulin concentration, with a calculated EC50 of 18.4 ± 1.1 nM. Conclusions: A simple and low-cost bioassay is proposed to quantify glucose consumption in 3T3-L1 adipose cells. Biological Sciences; Cell biology; Cell Culture; Cell Differentiation; Metabolism; Biological Assay; bioassay; In Vitro Techniques; Adipocytes; 3T3-L1 Cells; Glucose consumption; Glucose uptake; Insulin; Insulin action; Insulin response. © 2020 The Authorseng
dc.format.mimetypeapplication/pdf
dc.identifier.doihttps://doi.org/10.1016/j.heliyon.2020.e03422
dc.identifier.issn24058440
dc.identifier.urihttps://repository.urosario.edu.co/handle/10336/24191
dc.language.isoengspa
dc.publisherElsevier Ltdspa
dc.relation.citationIssueNo. 2
dc.relation.citationTitleHeliyon
dc.relation.citationVolumeVol. 6
dc.relation.ispartofHeliyon, ISSN:24058440, Vol.6, No.2 (2020)spa
dc.relation.urihttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85079861292&doi=10.1016%2fj.heliyon.2020.e03422&partnerID=40&md5=13877de2927975e4a49c3a52dec0a43fspa
dc.rights.accesRightsinfo:eu-repo/semantics/openAccess
dc.rights.accesoAbierto (Texto Completo)spa
dc.source.instnameinstname:Universidad del Rosariospa
dc.source.reponamereponame:Repositorio Institucional EdocURspa
dc.subject.keyword3T3-L1 cellsspa
dc.subject.keywordAdipocytesspa
dc.subject.keywordBioassayspa
dc.subject.keywordBiological assayspa
dc.subject.keywordBiological sciencesspa
dc.subject.keywordCell biologyspa
dc.subject.keywordCell culturespa
dc.subject.keywordCell differentiationspa
dc.subject.keywordGlucose consumptionspa
dc.subject.keywordGlucose uptakespa
dc.subject.keywordIn vitro techniquesspa
dc.subject.keywordInsulinspa
dc.subject.keywordInsulin actionspa
dc.subject.keywordInsulin responsespa
dc.subject.keywordMetabolismspa
dc.titleA colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cellsspa
dc.typearticleeng
dc.type.hasVersioninfo:eu-repo/semantics/publishedVersion
dc.type.spaArtículospa
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