title: Characterization of microglia induced from mouse embryonic stem cells and their migration into the brain parenchyma.
abstract: We derived microglia from mouse embryonic stem cells (ES cells) at very high density. Using the markers Mac1(+)/CD45(low) and Mac1(+)/CD45(high) to define microglia and macrophages, respectively, we show that Mac1(+) cells are induced by GM-CSF stimulation following neuronal differentiation of mouse ES cells using a five-step method. CD45(low) expression was high and CD45(high) expression was low on induced cells. We used a density gradient method to obtain a large amount of microglia-like cells, approximately 90% of Mac1(+) cells. Microglia-like cells expressed MHC class I, class II, CD40, CD80, CD86, and IFN-gammaR. The expression level of these molecules on microglia-like cells was barely enhanced by IFN-gamma. Intravenously transferred GFP(+) microglia derived from GFP(+) ES cells selectively accumulated in brain but not in peripheral tissues such as spleen and lymph node. GFP(+) cells were detected mainly in corpus callosum and hippocampus but were rarely seen in cerebral cortex, where Iba1, another marker of microglia, is primarily expressed. Furthermore, both GFP(+) and Iba1(+) cells exhibited a ramified morphology characteristic of mature microglia. These studies suggest that ES cell-derived microglia-like cells obtained using our protocol are functional and migrate selectively into the brain but not into peripheral tissues after intravenous transplantation.
authors.affiliation: Department of Neurosurgery, Kochi Medical School, Kochi University, Kohasu, Okoh-cho, Nankoku, Kochi 783-8505, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsuchiya
authors.forename: Kae Chang
authors.initials: KC
authors.lastname: Park
authors.forename: Shinichi
authors.initials: S
authors.lastname: Toyonaga
authors.forename: Shoko M
authors.initials: SM
authors.lastname: Yamada
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Nakabayashi
authors.forename: Eiichi
authors.initials: E
authors.lastname: Nakai
authors.forename: Naoki
authors.initials: N
authors.lastname: Ikawa
authors.forename: Masato
authors.initials: M
authors.lastname: Furuya
authors.forename: Akira
authors.initials: A
authors.lastname: Tominaga
authors.forename: Keiji
authors.initials: K
authors.lastname: Shimizu
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D000954:Antigens, Surface / Q000096:biosynthesis; D001921:Brain / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D002454:Cell Differentiation / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D004622:Embryo, Mammalian / Q000166:cytology*; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis; D000949:Histocompatibility Antigens Class II / Q000096:biosynthesis; D007275:Injections, Intravenous; D008264:Macrophages / Q000166:cytology / Q000276:immunology / Q000637:transplantation; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D017628:Microglia / Q000166:cytology* / Q000276:immunology / Q000637:transplantation; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology
chemical_list: D015703:Antigens, CD; D000954:Antigens, Surface; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D049452:Green Fluorescent Proteins