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Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker

Eom, Jennifer; Feisst, Vaughan; Ranjard, Louis; Loomes, Kerry; Damani, Tanvi; Jackson-Patel, Victoria; Locke, Michelle; Sheppard, Hilary; Narayan, Pritika; Dunbar, P. Rod

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Several dyes are currently available for use in detecting differentiation of mesenchymal cells into adipocytes. Dyes, such as Oil Red O, are cheap, easy to use and widely utilized by laboratories analyzing the adipogenic potential of mesenchymal cells. However, they are not specific to changes in gene transcription. We have developed a gene-specific differentiation assay to analyze when a mesenchymal cell has switched its fate to an adipogenic lineage. Immuno-labelling against fatty acid...[Show more]

dc.contributor.authorEom, Jennifer
dc.contributor.authorFeisst, Vaughan
dc.contributor.authorRanjard, Louis
dc.contributor.authorLoomes, Kerry
dc.contributor.authorDamani, Tanvi
dc.contributor.authorJackson-Patel, Victoria
dc.contributor.authorLocke, Michelle
dc.contributor.authorSheppard, Hilary
dc.contributor.authorNarayan, Pritika
dc.contributor.authorDunbar, P. Rod
dc.date.accessioned2019-10-10T02:38:48Z
dc.date.available2019-10-10T02:38:48Z
dc.identifier.citationEom, J., Feisst, V., Ranjard, L., Loomes, K., Damani, T., Jackson-Patel, V., Locke, M., Sheppard, H., Narayan, P., Dunbar, P.R. Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker. J. Vis. Exp. (133), e57153, doi:10.3791/57153 (2018).
dc.identifier.issn1940-087X
dc.identifier.urihttp://hdl.handle.net/1885/173623
dc.description.abstractSeveral dyes are currently available for use in detecting differentiation of mesenchymal cells into adipocytes. Dyes, such as Oil Red O, are cheap, easy to use and widely utilized by laboratories analyzing the adipogenic potential of mesenchymal cells. However, they are not specific to changes in gene transcription. We have developed a gene-specific differentiation assay to analyze when a mesenchymal cell has switched its fate to an adipogenic lineage. Immuno-labelling against fatty acid binding protein-4 (FABP4), a lineage-specific marker of adipogenic differentiation, enabled visualization and quantification of differentiated cells. The ability to quantify adipogenic differentiation potential of mesenchymal cells in a 96 well microplate format has promising implications for a number of applications. Hundreds of clinical trials involve the use of adult mesenchymal stromal cells and it is currently difficult to correlate therapeutic outcomes within and especially between such clinical trials. This simple high-throughput FABP4 assay provides a quantitative assay for assessing the differentiation potential of patient-derived cells and is a robust tool for comparing different isolation and expansion methods. This is particularly important given the increasing recognition of the heterogeneity of the cells being administered to patients in mesenchymal cell products. The assay also has potential utility in high throughput drug screening, particularly in obesity and pre-diabetes research.
dc.description.sponsorshipWe are also grateful to the University of Auckland, Faculty of Medical and Health Sciences Performance Based Research Fund for funding the cost of videoing and editing for submission of this article.
dc.format.mimetypeapplication/pdf
dc.language.isoen_AU
dc.publisherJournal of Visualized Experiments
dc.rights© 2018
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/
dc.sourceJournal of Visualized Experiments
dc.subjectAdipose derived stromal cells
dc.subjectdifferentiation
dc.subjectadipogenesis
dc.subjectimaging
dc.subjectimmunocytochemistry
dc.subjectimmunofluorescence
dc.subjectquantification
dc.subjecthigh content screening
dc.titleVisualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker
dc.typeJournal article
local.description.notesImported from ARIES
local.identifier.citationvolume133
dc.date.issued2018-03-31
local.identifier.absfor060104 - Cell Metabolism
local.identifier.ariespublicationu4485658xPUB2156
local.publisher.urlhttps://www.jove.com
local.type.statusPublished Version
local.contributor.affiliationEom, Jennifer, University of Auckland
local.contributor.affiliationFeisst, Vaughan, University of Auckland
local.contributor.affiliationRanjard, Louis, College of Science, ANU
local.contributor.affiliationLoomes, Kerry, University of Auckland
local.contributor.affiliationDamani, Tanvi, University of Auckland
local.contributor.affiliationJackson-Patel, Victoria, University of Auckland
local.contributor.affiliationLocke, Michelle, University of Auckland
local.contributor.affiliationSheppard, Hilary, University of Auckland
local.contributor.affiliationNarayan, Pritika, University of Auckland
local.contributor.affiliationDunbar, P Rod, University of Auckland
local.bibliographicCitation.startpage1
local.bibliographicCitation.lastpage8
local.identifier.doi10.3791/57153
local.identifier.absseo970106 - Expanding Knowledge in the Biological Sciences
dc.date.updated2019-04-21T08:30:33Z
local.identifier.scopusID2-s2.0-85049910632
local.identifier.thomsonID000443329500034
dcterms.accessRightsOpen Access
dc.provenanceArticle states Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License
dc.rights.licenseCreative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported
CollectionsANU Research Publications

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