Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/133431
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Type: | Journal article |
Title: | A CX3CR1 reporter hESC line facilitates integrative analysis of in-vitro-derived microglia and improved microglia identity upon neuron-glia co-culture |
Author: | Grubman, A. Vandekolk, T.H. Schröder, J. Sun, G. Hatwell-Humble, J. Chan, J. Oksanen, M. Lehtonen, S. Hunt, C. Koistinaho, J.E. Nilsson, S.K. Haynes, J.M. Pouton, C.W. Polo, J.M. |
Citation: | Stem Cell Reports, 2020; 14(6):1018-1032 |
Publisher: | Elsevier |
Issue Date: | 2020 |
ISSN: | 2213-6711 2213-6711 |
Statement of Responsibility: | Alexandra Grubman, Teresa H.Vandekolk, Jan Schröder, Guizhi Sun, Jessica Hatwell-Humble .. et al. |
Abstract: | Multiple 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. |
Keywords: | Microglia; in-vitro-derived microglia; microglia genetic reporter; PSC differentiation; synapse internalization; RNA-seq; integrative molecular analysis |
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/ |
DOI: | 10.1016/j.stemcr.2020.04.007 |
Grant ID: | http://purl.org/au-research/grants/nhmrc/1097461 http://purl.org/au-research/grants/nhmrc/1105786 http://purl.org/au-research/grants/nhmrc/109937 |
Published version: | http://dx.doi.org/10.1016/j.stemcr.2020.04.007 |
Appears in Collections: | Medical Sciences publications |
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hdl_133431.pdf | Published version | 3.97 MB | Adobe PDF | View/Open |
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