title: Development of a culture system to induce microglia-like cells from haematopoietic cells.
abstract: OBJECTIVE
  Microglia are the resident immune cells in the central nervous system, originating from haematopoietic-derived myeloid cells. A microglial cell is a double-edged sword, which has both pro-inflammatory and anti-inflammatory functions. Although understanding the role of microglia in pathological conditions has become increasingly important, histopathology has been the only way to investigate microglia in human diseases.
  
  
  METHODS
  To enable the study of microglial cells in vitro, we here establish a culture system to induce microglia-like cells from haematopoietic cells by coculture with astrocytes. The characteristics of microglia-like cells were analysed by flow cytometry and functional assay.
  
  
  RESULTS
  We show that triggering receptor expressing on myeloid cells-2-expressing microglia-like cells could be induced from lineage negative cells or monocytes by coculture with astrocytes. Microglia-like cells exhibited lower expression of CD45 and MHC class II than macrophages, a characteristic similar to brain microglia. When introduced into brain slice cultures, these microglia-like cells changed their morphology to a ramified shape on the first day of the culture. Moreover, we demonstrated that microglia-like cells could be induced from human monocytes by coculture with astrocytes. Finally, we showed that interleukin 34 was an important factor in the induction of microglia-like cells from haematopoietic cells in addition to cell-cell contact with astrocytes. Purified microglia-like cells were suitable for further culture and functional analyses.
  
  
  CONCLUSIONS
  Development of in vitro induction system for microglia will further promote the study of human microglial cells under pathological conditions as well as aid in the screening of drugs to target microglial cells.
authors.affiliation: Department of Immunology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo, Japan; Department of Neurology and Neurobiology of Aging, Kanazawa University Graduate School of Medical Science, Kanazawa, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Noto
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sakuma
authors.forename: Kazuya
authors.initials: K
authors.lastname: Takahashi
authors.forename: Reiko
authors.initials: R
authors.lastname: Saika
authors.forename: Ryoko
authors.initials: R
authors.lastname: Saga
authors.forename: Masahito
authors.initials: M
authors.lastname: Yamada
authors.forename: Takashi
authors.initials: T
authors.lastname: Yamamura
authors.forename: Sachiko
authors.initials: S
authors.lastname: Miyake
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000502:physiology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002540:Cerebral Cortex / Q000166:cytology; D018920:Coculture Techniques / Q000379:methods*; D005260:Female; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D007378:Interleukins / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D017628:Microglia / Q000166:cytology* / Q000502:physiology*; D009000:Monocytes / Q000187:drug effects / Q000502:physiology*; D013116:Spinal Cord / Q000166:cytology
chemical_list: D007378:Interleukins; C552364:interleukin-34, mouse
keywords: astrocytes; haematopoietic cells; interleukin 34; microglia; monocytes; triggering receptor expressing on myeloid cells-2 (TREM2)
references.citation: Cell Death Differ. 2010 Dec;17(12):1917-27
references.pmid: 20489731
references.citation: J Neurochem. 2009 Jul;110(1):284-94
references.pmid: 19457124
references.citation: PLoS Med. 2007 Apr;4(4):e124
references.pmid: 17425404
references.citation: Science. 2010 Nov 5;330(6005):841-5
references.pmid: 20966214
references.citation: FASEB J. 2004 Jun;18(9):998-1000
references.pmid: 15084516
references.citation: J Neurosci. 2013 Feb 6;33(6):2481-93
references.pmid: 23392676
references.citation: Eur J Neurosci. 2004 Nov;20(10):2617-28
references.pmid: 15548205
references.citation: Glia. 1996 Sep;18(1):1-10
references.pmid: 8891687
references.citation: BMC Immunol. 2002 Oct 24;3:15
references.pmid: 12398794
references.citation: Glia. 1998 Feb;22(2):180-8
references.pmid: 9537838
references.citation: Brain Res Brain Res Rev. 2005 Apr;48(2):196-206
references.pmid: 15850658
references.citation: J Exp Med. 2005 Feb 21;201(4):647-57
references.pmid: 15728241
references.citation: J Immunol. 1995 May 1;154(9):4309-21
references.pmid: 7722289
references.citation: Nat Neurosci. 2007 Dec;10(12):1544-53
references.pmid: 18026096
references.citation: Eur J Neurosci. 2010 Apr;31(7):1155-63
references.pmid: 20345919
references.citation: Nat Rev Immunol. 2003 Jun;3(6):445-53
references.pmid: 12776204
references.citation: Am J Pathol. 2011 Oct;179(4):2016-27
references.pmid: 21872563
references.citation: Annu Rev Immunol. 2009;27:119-45
references.pmid: 19302036
references.citation: Nat Med. 2011 Jan;17(1):64-70
references.pmid: 21131957
references.citation: J Inflamm (Lond). 2012 Apr 02;9(1):12
references.pmid: 22471998
references.citation: Brain Res Rev. 2007 Jun;54(2):274-85
references.pmid: 17383006
references.citation: J Leukoc Biol. 2010 Sep;88(3):495-505
references.pmid: 20504948
references.citation: Proc Natl Acad Sci U S A. 2001 May 22;98(11):6295-300
references.pmid: 11371643
references.citation: Nat Neurosci. 2007 Dec;10(12):1538-43
references.pmid: 18026097
references.citation: J Neurochem. 2002 Dec;83(6):1309-20
references.pmid: 12472885
references.citation: J Leukoc Biol. 2004 Mar;75(3):388-97
references.pmid: 14612429
references.citation: Cell. 2010 May 28;141(5):775-85
references.pmid: 20510925
references.citation: J Leukoc Biol. 2009 Mar;85(3):352-70
references.pmid: 19028958
references.citation: Nat Immunol. 2012 Jun 24;13(8):753-60
references.pmid: 22729249
references.citation: Nat Med. 2001 Dec;7(12):1356-61
references.pmid: 11726978
references.citation: Acta Neuropathol. 2009 Aug;118(2):219-33
references.pmid: 19533155
references.citation: Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7438-42
references.pmid: 1651506
references.citation: J Exp Med. 2008 Sep 29;205(10):2319-37
references.pmid: 18779347
references.citation: Science. 1988 Jan 15;239(4837):290-2
references.pmid: 3276004
references.citation: Science. 2009 Jul 31;325(5940):612-6
references.pmid: 19644120
references.citation: Eur J Neurosci. 2001 Aug;14(3):463-73
references.pmid: 11553296
references.citation: Nat Med. 2004 Feb;10(2):117-8; author reply 118-9
references.pmid: 14760408
references.citation: Annu Rev Immunol. 2009;27:669-92
references.pmid: 19132917
references.citation: ASN Neuro. 2010 Jul 12;2(3):e00037
references.pmid: 20640189
references.citation: J Cell Biol. 2009 Jun 1;185(5):779-86
references.pmid: 19487453
references.citation: Science. 2008 May 9;320(5877):807-11
references.pmid: 18467591
references.citation: Curr Opin Neurobiol. 2008 Oct;18(5):488-94
references.pmid: 18840525
references.citation: J Neuroimmunol. 2007 Mar;184(1-2):92-9
references.pmid: 17239445