A novel in vitro human microglia model: Characterization of human monocyte-derived microglia

Etemad, Samar, Zamin, Rasheeda Mohd, Ruitenberg, Marc J. and Filgueira, Luis (2012) A novel in vitro human microglia model: Characterization of human monocyte-derived microglia. Journal of Neuroscience Methods, 209 1: 79-89.


Author Etemad, Samar
Zamin, Rasheeda Mohd
Ruitenberg, Marc J.
Filgueira, Luis
Title A novel in vitro human microglia model: Characterization of human monocyte-derived microglia
Journal name Journal of Neuroscience Methods   Check publisher's open access policy
ISSN 0165-0270
1872-678X
Publication date 2012-07
Sub-type Article (original research)
DOI 10.1016/j.jneumeth.2012.05.025
Volume 209
Issue 1
Start page 79
End page 89
Total pages 11
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2013
Language eng
Formatted abstract Microglia are the innate immune cells of the central nervous system. They help maintaining physiological homeostasis and contribute significantly to inflammatory responses in the course of infection, injury and degenerative processes. To date, there is no standardized simple model available to investigate the biology of humanmicroglia. The aim of this study was to establish a new humanmicroglia model. For that purpose, human peripheral blood monocytes were cultured in serum free medium in the presence of M-CSF, GM-CSF, NGF and CCL2 to generate monocyte-derivedmicroglia (M-MG). M-MG were clearly different in morphology, phenotype and function from freshly isolated monocytes, cultured monocytes in the absence of the cytokines and monocyte-derived dendritic cells (M-DC) cultured in the presence of GM-CSF and IL-4. M-MG acquired a ramified morphology with primary and secondary processes. M-MG displayed a comparable phenotype to the humanmicroglia cell line HMC3, expressing very low levels of CD45, CD14 and HLA-DR, CD11b and CD11c; and undetectable levels of CD40, CD80 and CD83, and a distinct pattern of chemokine receptors (positive for CCR1, CCR2, CCR4, CCR5, CXCR1, CXCR3, CX3CR1; negative for CCR6 and CCR7). In comparison with M-DC, M-MG displayed lower T-lymphocyte stimulatory capacity, as well as lower phagocytosis activity. The described protocol for the generation of humanmonocyte-derivedmicroglia is feasible, well standardized and reliable, as it uses well defined culture medium and recombinant cytokines, but no serum or conditioned medium. This protocol will certainly be very helpful for future studies investigating the biology and pathology of humanmicroglia.
Keyword Microglia
Human peripheral blood monocytes
Serum-free medium
Flow cytometry
Chemokine receptors
M-Csf
Gm-Csf
Ccl2
Ngf
Phagocytosis
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2013 Collection
School of Biomedical Sciences Publications
 
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