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dc.contributor.authorChen, Jian-Hua
dc.contributor.authorGoh, Kim Jee
dc.contributor.authorRocha, Nuno
dc.contributor.authorGroeneveld, Matthijs P
dc.contributor.authorMinic, Marina
dc.contributor.authorBarrett, Timothy G
dc.contributor.authorSavage, David
dc.contributor.authorSemple, Robert
dc.date.accessioned2018-05-25T11:18:17Z
dc.date.available2018-05-25T11:18:17Z
dc.date.issued2017-12-19
dc.identifier.issn1754-8403
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/276194
dc.description.abstractAdipose tissue is the primary tissue affected in most single gene forms of severe insulin resistance, and growing evidence has implicated it as a site at which many risk alleles for insulin resistance identified in population-wide studies might exert their effect. There is thus increasing need for human adipocyte models in which to interrogate the function of known and emerging genetic risk variants. However, primary adipocyte cultures, existing immortalised cell lines and stem-cell based models all have significant biological or practical limitations. In an attempt to widen the repertoire of human cell models in which to study adipocyte-autonomous effects of relevant human genetic variants, we have undertaken direct reprogramming of skin fibroblasts to adipocyte-like cells by employing an inducible recombinant lentivirus overexpressing the master adipogenic transcription factor PPARγ2. Doxycycline-driven expression of PPARγ2 and adipogenic culture conditions converted dermal fibroblasts into triglyceride-laden cells within days. The resulting cells recapitulated most of the crucial aspects of adipocyte biology in vivo, including the expression of mature adipocyte markers, secreted high levels of the adipokine adiponectin, and underwent lipolysis when treated with isoproterenol/3-isobutyl-1-methylxanthine (IBMX). They did not, however, exhibit insulin-inducible glucose uptake, and withdrawal of doxycycline produced rapid delipidation and loss of adipogenic markers. This protocol was applied successfully to a panel of skin cells from individuals with monogenic severe insulin resistance; however, surprisingly, even cell lines harbouring mutations causing severe, generalised lipodystrophy accumulated large lipid droplets and induced adipocyte-specific genes. The direct reprogramming protocol of human dermal fibroblasts to adipocyte-like cells we established is simple, fast and efficient, and has the potential to generate cells which can serve as a tool to address some, though not all, aspects of adipocyte function in the presence of endogenous disease-causing mutations.
dc.format.mediumElectronic
dc.languageeng
dc.publisherThe Company of Biologists
dc.rightsAttribution 4.0 International
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectAdipocytes
dc.subjectFibroblasts
dc.subjectStem Cells
dc.subjectDermis
dc.subjectHumans
dc.subjectMetabolic Diseases
dc.subjectPPAR gamma
dc.subjectMutation
dc.subjectModels, Biological
dc.subjectCellular Reprogramming
dc.titleEvaluation of human dermal fibroblasts directly reprogrammed to adipocyte-like cells as a metabolic disease model.
dc.typeArticle
prism.endingPage1420
prism.issueIdentifier12
prism.publicationDate2017
prism.publicationNameDis Model Mech
prism.startingPage1411
prism.volume10
dc.identifier.doi10.17863/CAM.23477
dcterms.dateAccepted2017-09-29
rioxxterms.versionofrecord10.1242/dmm.030981
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
rioxxterms.licenseref.startdate2017-12-19
dc.contributor.orcidGroeneveld, Matthijs P [0000-0002-3016-4853]
dc.contributor.orcidSavage, David [0000-0002-7857-7032]
dc.contributor.orcidSemple, Robert [0000-0001-6539-3069]
dc.identifier.eissn1754-8411
rioxxterms.typeJournal Article/Review
pubs.funder-project-idWellcome Trust (098498/Z/12/Z)
pubs.funder-project-idWellcome Trust (100574/Z/12/Z)
pubs.funder-project-idMedical Research Council (MC_UU_12012/5)
pubs.funder-project-idWellcome Trust (095515/Z/11/Z)
cam.issuedOnline2017-01-01


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International