Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker
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Eom, Jennifer; Feisst, Vaughan; Ranjard, Louis; Loomes, Kerry; Damani, Tanvi; Jackson-Patel, Victoria; Locke, Michelle; Sheppard, Hilary; Narayan, Pritika; Dunbar, P. Rod
Description
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.author | Eom, Jennifer | |
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dc.contributor.author | Feisst, Vaughan | |
dc.contributor.author | Ranjard, Louis | |
dc.contributor.author | Loomes, Kerry | |
dc.contributor.author | Damani, Tanvi | |
dc.contributor.author | Jackson-Patel, Victoria | |
dc.contributor.author | Locke, Michelle | |
dc.contributor.author | Sheppard, Hilary | |
dc.contributor.author | Narayan, Pritika | |
dc.contributor.author | Dunbar, P. Rod | |
dc.date.accessioned | 2019-10-10T02:38:48Z | |
dc.date.available | 2019-10-10T02:38:48Z | |
dc.identifier.citation | Eom, 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.issn | 1940-087X | |
dc.identifier.uri | http://hdl.handle.net/1885/173623 | |
dc.description.abstract | 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 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.sponsorship | We 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.mimetype | application/pdf | |
dc.language.iso | en_AU | |
dc.publisher | Journal of Visualized Experiments | |
dc.rights | © 2018 | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/3.0/ | |
dc.source | Journal of Visualized Experiments | |
dc.subject | Adipose derived stromal cells | |
dc.subject | differentiation | |
dc.subject | adipogenesis | |
dc.subject | imaging | |
dc.subject | immunocytochemistry | |
dc.subject | immunofluorescence | |
dc.subject | quantification | |
dc.subject | high content screening | |
dc.title | Visualization and Quantification of Mesenchymal Cell Adipogenic Differentiation Potential with a Lineage Specific Marker | |
dc.type | Journal article | |
local.description.notes | Imported from ARIES | |
local.identifier.citationvolume | 133 | |
dc.date.issued | 2018-03-31 | |
local.identifier.absfor | 060104 - Cell Metabolism | |
local.identifier.ariespublication | u4485658xPUB2156 | |
local.publisher.url | https://www.jove.com | |
local.type.status | Published Version | |
local.contributor.affiliation | Eom, Jennifer, University of Auckland | |
local.contributor.affiliation | Feisst, Vaughan, University of Auckland | |
local.contributor.affiliation | Ranjard, Louis, College of Science, ANU | |
local.contributor.affiliation | Loomes, Kerry, University of Auckland | |
local.contributor.affiliation | Damani, Tanvi, University of Auckland | |
local.contributor.affiliation | Jackson-Patel, Victoria, University of Auckland | |
local.contributor.affiliation | Locke, Michelle, University of Auckland | |
local.contributor.affiliation | Sheppard, Hilary, University of Auckland | |
local.contributor.affiliation | Narayan, Pritika, University of Auckland | |
local.contributor.affiliation | Dunbar, P Rod, University of Auckland | |
local.bibliographicCitation.startpage | 1 | |
local.bibliographicCitation.lastpage | 8 | |
local.identifier.doi | 10.3791/57153 | |
local.identifier.absseo | 970106 - Expanding Knowledge in the Biological Sciences | |
dc.date.updated | 2019-04-21T08:30:33Z | |
local.identifier.scopusID | 2-s2.0-85049910632 | |
local.identifier.thomsonID | 000443329500034 | |
dcterms.accessRights | Open Access | |
dc.provenance | Article states Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License | |
dc.rights.license | Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported | |
Collections | ANU Research Publications |
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