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

Introduction: 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...

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Autores:
Tipo de recurso:
Fecha de publicación:
2020
Institución:
Universidad del Rosario
Repositorio:
Repositorio EdocUR - U. Rosario
Idioma:
eng
OAI Identifier:
oai:repository.urosario.edu.co:10336/24191
Acceso en línea:
https://doi.org/10.1016/j.heliyon.2020.e03422
https://repository.urosario.edu.co/handle/10336/24191
Palabra clave:
3T3-L1 cells
Adipocytes
Bioassay
Biological assay
Biological sciences
Cell biology
Cell culture
Cell differentiation
Glucose consumption
Glucose uptake
In vitro techniques
Insulin
Insulin action
Insulin response
Metabolism
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License
Abierto (Texto Completo)
id EDOCUR2_865fc8b8aeb8992268a3f7d28abba731
oai_identifier_str oai:repository.urosario.edu.co:10336/24191
network_acronym_str EDOCUR2
network_name_str Repositorio EdocUR - U. Rosario
repository_id_str
spelling 49ae12f7-1bf7-4a42-814f-35c67460de117a1d2cc2-a3a5-47c9-9e65-f6d57246bbb27983198160035ff6196-a183-4161-9e6c-109909cc15092020-05-26T00:09:56Z2020-05-26T00:09:56Z2020Introduction: 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 Authorsapplication/pdfhttps://doi.org/10.1016/j.heliyon.2020.e0342224058440https://repository.urosario.edu.co/handle/10336/24191engElsevier LtdNo. 2HeliyonVol. 6Heliyon, ISSN:24058440, Vol.6, No.2 (2020)https://www.scopus.com/inward/record.uri?eid=2-s2.0-85079861292&doi=10.1016%2fj.heliyon.2020.e03422&partnerID=40&md5=13877de2927975e4a49c3a52dec0a43fAbierto (Texto Completo)http://purl.org/coar/access_right/c_abf2instname:Universidad del Rosarioreponame:Repositorio Institucional EdocUR3T3-L1 cellsAdipocytesBioassayBiological assayBiological sciencesCell biologyCell cultureCell differentiationGlucose consumptionGlucose uptakeIn vitro techniquesInsulinInsulin actionInsulin responseMetabolismA colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cellsarticleArtículohttp://purl.org/coar/version/c_970fb48d4fbd8a85http://purl.org/coar/resource_type/c_6501Diaz, Paola A. RiveraCamargo, Doris E. GómezOndo Méndez, Alejandro OyonoGómez-Alegría, Claudio J.10336/24191oai:repository.urosario.edu.co:10336/241912022-05-02 07:37:16.869907https://repository.urosario.edu.coRepositorio institucional EdocURedocur@urosario.edu.co
dc.title.spa.fl_str_mv A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
title A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
spellingShingle A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
3T3-L1 cells
Adipocytes
Bioassay
Biological assay
Biological sciences
Cell biology
Cell culture
Cell differentiation
Glucose consumption
Glucose uptake
In vitro techniques
Insulin
Insulin action
Insulin response
Metabolism
title_short A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
title_full A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
title_fullStr A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
title_full_unstemmed A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
title_sort A colorimetric bioassay for quantitation of both basal and insulin-induced glucose consumption in 3T3-L1 adipose cells
dc.subject.keyword.spa.fl_str_mv 3T3-L1 cells
Adipocytes
Bioassay
Biological assay
Biological sciences
Cell biology
Cell culture
Cell differentiation
Glucose consumption
Glucose uptake
In vitro techniques
Insulin
Insulin action
Insulin response
Metabolism
topic 3T3-L1 cells
Adipocytes
Bioassay
Biological assay
Biological sciences
Cell biology
Cell culture
Cell differentiation
Glucose consumption
Glucose uptake
In vitro techniques
Insulin
Insulin action
Insulin response
Metabolism
description Introduction: 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 Authors
publishDate 2020
dc.date.accessioned.none.fl_str_mv 2020-05-26T00:09:56Z
dc.date.available.none.fl_str_mv 2020-05-26T00:09:56Z
dc.date.created.spa.fl_str_mv 2020
dc.type.eng.fl_str_mv article
dc.type.coarversion.fl_str_mv http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.coar.fl_str_mv http://purl.org/coar/resource_type/c_6501
dc.type.spa.spa.fl_str_mv Artículo
dc.identifier.doi.none.fl_str_mv https://doi.org/10.1016/j.heliyon.2020.e03422
dc.identifier.issn.none.fl_str_mv 24058440
dc.identifier.uri.none.fl_str_mv https://repository.urosario.edu.co/handle/10336/24191
url https://doi.org/10.1016/j.heliyon.2020.e03422
https://repository.urosario.edu.co/handle/10336/24191
identifier_str_mv 24058440
dc.language.iso.spa.fl_str_mv eng
language eng
dc.relation.citationIssue.none.fl_str_mv No. 2
dc.relation.citationTitle.none.fl_str_mv Heliyon
dc.relation.citationVolume.none.fl_str_mv Vol. 6
dc.relation.ispartof.spa.fl_str_mv Heliyon, ISSN:24058440, Vol.6, No.2 (2020)
dc.relation.uri.spa.fl_str_mv https://www.scopus.com/inward/record.uri?eid=2-s2.0-85079861292&doi=10.1016%2fj.heliyon.2020.e03422&partnerID=40&md5=13877de2927975e4a49c3a52dec0a43f
dc.rights.coar.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.rights.acceso.spa.fl_str_mv Abierto (Texto Completo)
rights_invalid_str_mv Abierto (Texto Completo)
http://purl.org/coar/access_right/c_abf2
dc.format.mimetype.none.fl_str_mv application/pdf
dc.publisher.spa.fl_str_mv Elsevier Ltd
institution Universidad del Rosario
dc.source.instname.spa.fl_str_mv instname:Universidad del Rosario
dc.source.reponame.spa.fl_str_mv reponame:Repositorio Institucional EdocUR
repository.name.fl_str_mv Repositorio institucional EdocUR
repository.mail.fl_str_mv edocur@urosario.edu.co
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