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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.

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  • Authors+Show Affiliations

    ,

    Department of Neurosurgery, Kochi Medical School, Kochi University, Kohasu, Okoh-cho, Nankoku, Kochi 783-8505, Japan.

    , , , , , , , ,

    Source

    Journal of neuroimmunology 160:1-2 2005 Mar pg 210-8

    MeSH

    Animals
    Antigens, CD
    Antigens, Surface
    Brain
    Cell Differentiation
    Cell Line
    Cell Movement
    Embryo, Mammalian
    Female
    Green Fluorescent Proteins
    Histocompatibility Antigens Class I
    Histocompatibility Antigens Class II
    Injections, Intravenous
    Macrophages
    Mice
    Mice, Inbred C57BL
    Mice, Transgenic
    Microglia
    Stem Cell Transplantation
    Stem Cells

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    15710475

    Citation

    Tsuchiya, Takahiro, et al. "Characterization of Microglia Induced From Mouse Embryonic Stem Cells and Their Migration Into the Brain Parenchyma." Journal of Neuroimmunology, vol. 160, no. 1-2, 2005, pp. 210-8.
    Tsuchiya T, Park KC, Toyonaga S, et al. Characterization of microglia induced from mouse embryonic stem cells and their migration into the brain parenchyma. J Neuroimmunol. 2005;160(1-2):210-8.
    Tsuchiya, T., Park, K. C., Toyonaga, S., Yamada, S. M., Nakabayashi, H., Nakai, E., ... Shimizu, K. (2005). Characterization of microglia induced from mouse embryonic stem cells and their migration into the brain parenchyma. Journal of Neuroimmunology, 160(1-2), pp. 210-8.
    Tsuchiya T, et al. Characterization of Microglia Induced From Mouse Embryonic Stem Cells and Their Migration Into the Brain Parenchyma. J Neuroimmunol. 2005;160(1-2):210-8. PubMed PMID: 15710475.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - Characterization of microglia induced from mouse embryonic stem cells and their migration into the brain parenchyma. AU - Tsuchiya,Takahiro, AU - Park,Kae Chang, AU - Toyonaga,Shinichi, AU - Yamada,Shoko M, AU - Nakabayashi,Hiromichi, AU - Nakai,Eiichi, AU - Ikawa,Naoki, AU - Furuya,Masato, AU - Tominaga,Akira, AU - Shimizu,Keiji, Y1 - 2004/12/24/ PY - 2004/09/13/received PY - 2004/10/25/revised PY - 2004/10/25/accepted PY - 2005/2/16/pubmed PY - 2005/4/29/medline PY - 2005/2/16/entrez SP - 210 EP - 8 JF - Journal of neuroimmunology JO - J. Neuroimmunol. VL - 160 IS - 1-2 N2 - 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. SN - 0165-5728 UR - https://www.unboundmedicine.com/medline/citation/15710475/Characterization_of_microglia_induced_from_mouse_embryonic_stem_cells_and_their_migration_into_the_brain_parenchyma_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-5728(04)00404-7 DB - PRIME DP - Unbound Medicine ER -