A CX3CR1 Reporter hESC Line Facilitates Integrative Analysis of In-Vitro-Derived Microglia and Improved Microglia Identity upon Neuron-Glia Co-culture.

Alexandra Grubman; Teresa H Vandekolk; Jan Schröder; Guizhi Sun; Jessica Hatwell-Humble; Jonathan Chan; Minna Oksanen; Sarka Lehtonen; Cameron Hunt; Jari E Koistinaho; Susan K Nilsson; John M Haynes; Colin W Pouton; Jose M Polo

Journal article

A CX3CR1 Reporter hESC Line Facilitates Integrative Analysis of In-Vitro-Derived Microglia and Improved Microglia Identity upon Neuron-Glia Co-culture.

Alexandra Grubman, Teresa H Vandekolk, Jan Schröder, Guizhi Sun, Jessica Hatwell-Humble, Jonathan Chan, Minna Oksanen, Sarka Lehtonen, Cameron Hunt, Jari E Koistinaho, Susan K Nilsson, John M Haynes, Colin W Pouton, Jose M Polo

Stem Cell Reports | Elsevier (Cell Press) | Published : 2020

DOI: 10.1016/j.stemcr.2020.04.007

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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 t..

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University of Melbourne Researchers

Susan Nilsson Author Clinical Pathology

Jan Schroeder Author Microbiology and Immunology

Grants

Awarded by NHMRC-ARC Dementia Research Development Fellowship

Awarded by Monash Joint Medicine-Pharmacy Grant

Awarded by Monash University International Network of Excellence Grant

Awarded by NHMRC

Awarded by ARC Future Fellowship

Funding Acknowledgements

A.G. is supported by an NHMRC -ARC Dementia Research Development Fellowship ( GNT1097461 ). A.G., J.M.P., C.W.P., and J.M.H. are supported by a Monash Joint Medicine-Pharmacy Grant ( JMP17-0669 ), a Monash University International Network of Excellence Grant ( NOE170089 ), and an NHMRC Grant APP1105786 , and C.W.P. and J.M.H. by NHMRC grant APP1099372 . A.G. was supported by the Dementia Australia Research Foundation and Yulgilbar Foundation Innovation Grant. G.S. and J C. were supported by the Yulgilbar Foundation . J.M.P. was supported by Sylvia and Charles Viertel Senior Medical Research Fellowship and ARC Future Fellowship ( FT180100674 ). J.E.K. was supported by the Sigrid Jusélius Foundation , Finland. S.K.N. is supported by an OCE CSIRO Science Leader Fellowship. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government .