A CX3CR1 reporter hESC line facilitates integrative analysis of in-vitro-derived microglia and improved microglia identity upon neuron-glia co-culture

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dc.contributor.authorGrubman, A.
dc.contributor.authorVandekolk, T.H.
dc.contributor.authorSchröder, J.
dc.contributor.authorSun, G.
dc.contributor.authorHatwell-Humble, J.
dc.contributor.authorChan, J.
dc.contributor.authorOksanen, M.
dc.contributor.authorLehtonen, S.
dc.contributor.authorHunt, C.
dc.contributor.authorKoistinaho, J.E.
dc.contributor.authorNilsson, S.K.
dc.contributor.authorHaynes, J.M.
dc.contributor.authorPouton, C.W.
dc.contributor.authorPolo, J.M.
dc.date.issued2020
dc.description.abstractMultiple protocols have been published for generation of iMGLs from hESCs/iPSCs. To date, there are no guides to assist researchers to determine the most appropriate methodology for microglial studies. To establish a framework to facilitate future microglial studies, we first performed a comparative transcriptional analysis between iMGLs derived using three published datasets, which allowed us to establish the baseline protocol that is most representative of bona fide human microglia. Secondly, using CRISPR to tag the classic microglial marker CX3CR1 with nanoluciferase and tdTomato, we generated and functionally validated a reporter ESC line. Finally, using this cell line, we demonstrated that co-culture of iMGL precursors with human glia and neurons enhanced transcriptional resemblance of iMGLs to ex vivo microglia. Together, our comprehensive molecular analysis and reporter cell line are a useful resource for neurobiologists seeking to use iMGLs for disease modeling and drug screening studies.
dc.description.statementofresponsibilityAlexandra Grubman, Teresa H.Vandekolk, Jan Schröder, Guizhi Sun, Jessica Hatwell-Humble .. et al.
dc.identifier.citationStem Cell Reports, 2020; 14(6):1018-1032
dc.identifier.doi10.1016/j.stemcr.2020.04.007
dc.identifier.issn2213-6711
dc.identifier.issn2213-6711
dc.identifier.orcidPolo, J.M. [0000-0002-2531-778X]
dc.identifier.urihttps://hdl.handle.net/2440/133431
dc.language.isoen
dc.publisherElsevier
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1097461
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1105786
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/109937
dc.rights© 2020 The Authors. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.source.urihttps://doi.org/10.1016/j.stemcr.2020.04.007
dc.subjectMicroglia; in-vitro-derived microglia; microglia genetic reporter; PSC differentiation; synapse internalization; RNA-seq; integrative molecular analysis
dc.subject.meshMicroglia
dc.subject.meshNeurons
dc.subject.meshCell Line
dc.subject.meshHumans
dc.subject.meshCoculture Techniques
dc.subject.meshCell Differentiation
dc.subject.meshGenes, Reporter
dc.subject.meshTranscriptome
dc.subject.meshHuman Embryonic Stem Cells
dc.subject.meshCX3C Chemokine Receptor 1
dc.titleA CX3CR1 reporter hESC line facilitates integrative analysis of in-vitro-derived microglia and improved microglia identity upon neuron-glia co-culture
dc.typeJournal article
pubs.publication-statusPublished

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