Murine iPSC-derived microglia and macrophage cell culture models recapitulate distinct phenotypical and functional properties of classical and alternative neuro-immune polarisation.
Quarta, Alessandra; Le Blon, Debbie; D'Aes, Tineet al.
2019 • In Brain, Behavior and Immunity, 82, p. 406-421
[en] The establishment and validation of reliable induced pluripotent stem cell (iPSC)-derived in vitro models to study microglia and monocyte/macrophage immune function holds great potential for fundamental and translational neuro-immunology research. In this study, we first demonstrate that ramified CX3CR1(+) iPSC-microglia (cultured within a neural environment) and round-shaped CX3CR1(-) iPSC-macrophages can easily be differentiated from newly established murine CX3CR1(eGFP/+)CCR2(RFP/+) iPSC lines. Furthermore, we show that obtained murine iPSC-microglia and iPSC-macrophages are distinct cell populations, even though iPSC-macrophages may upregulate CX3CR1 expression when cultured within a neural environment. Next, we characterized the phenotypical and functional properties of murine iPSC-microglia and iPSC-macrophages following classical and alternative immune polarisation. While iPSC-macrophages could easily be triggered to adopt a classically-activated or alternatively-activated phenotype following, respectively, lipopolysaccharide+interferon gamma or interleukin 13 (IL13) stimulation, iPSC-microglia and iPSC-macrophages cultured within a neural environment displayed a more moderate activation profile as characterised by the absence of MHCII expression upon classical immune polarisation and the absence of Ym1 expression upon alternative immune polarisation. Finally, extending our preceding in vivo studies, this striking phenotypical divergence was also observed for resident microglia and infiltrating monocytes within highly inflammatory cortical lesions in CX3CR1(eGFP/+)CCR2(RFP/+) mice subjected to middle cerebral arterial occlusion (MCAO) stroke and following IL13-mediated therapeutic intervention thereon. In conclusion, our study demonstrates that the applied murine iPSC-microglia and iPSC-macrophage culture models are able to recapitulate in vivo microglia and monocyte/macrophage ontogeny and corresponding phenotypical/functional properties upon classical and alternative immune polarisation, and therefore represent a valuable in vitro platform to further study and modulate microglia and (infiltrating) monocyte immune responses under neuro-inflammatory conditions within a neural environment.
Disciplines :
Life sciences: Multidisciplinary, general & others
Murine iPSC-derived microglia and macrophage cell culture models recapitulate distinct phenotypical and functional properties of classical and alternative neuro-immune polarisation.
Publication date :
2019
Journal title :
Brain, Behavior and Immunity
ISSN :
0889-1591
eISSN :
1090-2139
Publisher :
Elsevier, Atlanta, United States - Florida
Volume :
82
Pages :
406-421
Peer reviewed :
Peer Reviewed verified by ORBi
Commentary :
Copyright (c) 2019 The Authors. Published by Elsevier Inc. All rights reserved.
Abud, E.M., Ramirez, R.N., Martinez, E.S., Healy, L.M., Nguyen, C.H.H., Newman, S.A., Yeromin, A.V., Scarfone, V.M., Marsh, S.E., Fimbres, C., Caraway, C.A., Fote, G.M., Madany, A.M., Agrawal, A., Kayed, R., Gylys, K.H., Cahalan, M.D., Cummings, B.J., Antel, J.P., Mortazavi, A., Carson, M.J., Poon, W.W., Blurton-Jones, M., 2017. iPSC-Derived Human Microglia-like Cells to Study Neurological Diseases. Neuron 94, 278-293 e279.
Abutbul, S., Shapiro, J., Szaingurten-Solodkin, I., Levy, N., Carmy, Y., Baron, R., Jung, S., Monsonego, A., 2012. TGF-beta signaling through SMAD2/3 induces the quiescent microglial phenotype within the CNS environment. Glia 60, 1160-1171.
Ajami, B., Bennett, J.L., Krieger, C., McNagny, K.M., Rossi, F.M., 2011. Infiltrating monocytes trigger EAE progression, but do not contribute to the resident microglia pool. Nat Neurosci 14, 1142-1149.
Benjamini, Y., Hochberg, Y., 1995. Controlling the False Discovery Rate - a Practical and Powerful Approach to Multiple Testing. J R Stat Soc B 57, 289-300.
Bennett, M.L., Bennett, F.C., Liddelow, S.A., Ajami, B., Zamanian, J.L., Fernhoff, N.B., Mulinyawe, S.B., Bohlen, C.J., Adil, A., Tucker, A., Weissman, I.L., Chang, E.F., Li, G., Grant, G.A., Hayden Gephart, M.G., Barres, B.A., 2016. New tools for studying microglia in the mouse and human CNS. Proc Natl Acad Sci U S A 113, E1738-1746.
Bennett, F.C., Bennett, M.L., Yaqoob, F., Mulinyawe, S.B., Grant, G.A., Hayden Gephart, M., Plowey, E.D., Barres, B.A., 2018. A Combination of Ontogeny and CNS Environment Establishes Microglial Identity. Neuron 98, 1170-1183 e1178.
Beutner, C., Roy, K., Linnartz, B., Napoli, I., Neumann, H., 2010. Generation of microglial cells from mouse embryonic stem cells. Nat Protoc 5, 1481-1494.
Bohlen, C.J., Bennett, F.C., Tucker, A.F., Collins, H.Y., Mulinyawe, S.B., Barres, B.A., 2017. Diverse Requirements for Microglial Survival, Specification, and Function Revealed by Defined-Medium Cultures. Neuron 94, 759-773 e758.
Brownjohn, P.W., Smith, J., Solanki, R., Lohmann, E., Houlden, H., Hardy, J., Dietmann, S., Livesey, F.J., 2018. Functional Studies of Missense TREM2 Mutations in Human Stem Cell-Derived Microglia. Stem Cell Reports 10, 1294-1307.
Cardona, A.E., Pioro, E.P., Sasse, M.E., Kostenko, V., Cardona, S.M., Dijkstra, I.M., Huang, D., Kidd, G., Dombrowski, S., Dutta, R., Lee, J.C., Cook, D.N., Jung, S., Lira, S.A., Littman, D.R., Ransohoff, R.M., 2006. Control of microglial neurotoxicity by the fractalkine receptor. Nat Neurosci 9, 917-924.
Cherry, J.D., Olschowka, J.A., O'Banion, M.K., 2014. Neuroinflammation and M2 microglia: the good, the bad, and the inflamed. J Neuroinflammation 11, 98.
Claes, C., Van Den Daele, J., Boon, R., Schouteden, S., Colombo, A., Monasor, L.S., Fiers, M., Ordovas, L., Nami, F., Bohrmann, B., Tahirovic, S., De Strooper, B., Verfaillie, C.M., 2019. Human stem cell-derived monocytes and microglia-like cells reveal impaired amyloid plaque clearance upon heterozygous or homozygous loss of TREM2. Alzheimers Dement 15, 453-464.
Costa, R., Bergwerf, I., Santermans, E., De Vocht, N., Praet, J., Daans, J., Le Blon, D., Hoornaert, C., Reekmans, K., Hens, N., Goossens, H., Berneman, Z., Parolini, O., Alviano, F., Ponsaerts, P., 2015. Distinct in vitro properties of embryonic and extraembryonic fibroblast-like cells are reflected in their in vivo behavior following grafting in the adult mouse brain. Cell Transplant 24, 223-233.
De Vocht, N., Praet, J., Reekmans, K., Le Blon, D., Hoornaert, C., Daans, J., Berneman, Z., Van der Linden, A., Ponsaerts, P., 2013. Tackling the physiological barriers for successful mesenchymal stem cell transplantation into the central nervous system. Stem Cell Res Ther 4, 101.
DiSabato, D.J., Quan, N., Godbout, J.P., 2016. Neuroinflammation: the devil is in the details. J Neurochem 139 Suppl 2, 136-153.
Dooley, D., Lemmens, E., Vangansewinkel, T., Le Blon, D., Hoornaert, C., Ponsaerts, P., Hendrix, S., 2016. Cell-Based Delivery of Interleukin-13 Directs Alternative Activation of Macrophages Resulting in Improved Functional Outcome after Spinal Cord Injury. Stem Cell Reports 7, 1099-1115.
Douvaras, P., Sun, B., Wang, M., Kruglikov, I., Lallos, G., Zimmer, M., Terrenoire, C., Zhang, B., Gandy, S., Schadt, E., Freytes, D.O., Noggle, S., Fossati, V., 2017. Directed Differentiation of Human Pluripotent Stem Cells to Microglia. Stem Cell Reports 8, 1516-1524.
Etemad, S., Zamin, R.M., Ruitenberg, M.J., Filgueira, L., 2012. A novel in vitro human microglia model: characterization of human monocyte-derived microglia. J Neurosci Methods 209, 79-89.
Garcia-Reitboeck, P., Phillips, A., Piers, T.M., Villegas-Llerena, C., Butler, M., Mallach, A., Rodrigues, C., Arber, C.E., Heslegrave, A., Zetterberg, H., Neumann, H., Neame, S., Houlden, H., Hardy, J., Pocock, J.M., 2018. Human Induced Pluripotent Stem Cell-Derived Microglia-Like Cells Harboring TREM2 Missense Mutations Show Specific Deficits in Phagocytosis. Cell Rep 24, 2300-2311.
Ginhoux, F., Greter, M., Leboeuf, M., Nandi, S., See, P., Gokhan, S., Mehler, M.F., Conway, S.J., Ng, L.G., Stanley, E.R., Samokhvalov, I.M., Merad, M., 2010. Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 330, 841-845.
Ginhoux, F., Jung, S., 2014. Monocytes and macrophages: developmental pathways and tissue homeostasis. Nat Rev Immunol 14, 392-404.
Gosselin, D., Link, V.M., Romanoski, C.E., Fonseca, G.J., Eichenfield, D.Z., Spann, N.J., Stender, J.D., Chun, H.B., Garner, H., Geissmann, F., Glass, C.K., 2014. Environment drives selection and function of enhancers controlling tissue-specific macrophage identities. Cell 159, 1327-1340.
Guglielmetti, C., Le Blon, D., Santermans, E., Salas-Perdomo, A., Daans, J., De Vocht, N., Shah, D., Hoornaert, C., Praet, J., Peerlings, J., Kara, F., Bigot, C., Mai, Z., Goossens, H., Hens, N., Hendrix, S., Verhoye, M., Planas, A.M., Berneman, Z., van der Linden, A., Ponsaerts, P., 2016. Interleukin-13 immune gene therapy prevents CNS inflammation and demyelination via alternative activation of microglia and macrophages. Glia 64, 2181-2200.
Gupta, N., Shyamasundar, S., Patnala, R., Karthikeyan, A., Arumugam, T.V., Ling, E.A., Dheen, S.T., 2018. Recent progress in therapeutic strategies for microglia-mediated neuroinflammation in neuropathologies. Expert Opin Ther Targets 22, 765-781.
Haenseler, W., Sansom, S.N., Buchrieser, J., Newey, S.E., Moore, C.S., Nicholls, F.J., Chintawar, S., Schnell, C., Antel, J.P., Allen, N.D., Cader, M.Z., Wade-Martins, R., James, W.S., Cowley, S.A., 2017. A Highly Efficient Human Pluripotent Stem Cell Microglia Model Displays a Neuronal-Co-culture-Specific Expression Profile and Inflammatory Response. Stem Cell Reports 8, 1727-1742.
Hamzei Taj, S., Le Blon, D., Hoornaert, C., Daans, J., Quarta, A., Praet, J., Van der Linden, A., Ponsaerts, P., Hoehn, M., 2018. Targeted intracerebral delivery of the anti-inflammatory cytokine IL13 promotes alternative activation of both microglia and macrophages after stroke. J Neuroinflammation 15, 174.
Hinze, A., Stolzing, A., 2011. Differentiation of mouse bone marrow derived stem cells toward microglia-like cells. BMC Cell Biol 12, 35.
Hoeffel, G., Chen, J., Lavin, Y., Low, D., Almeida, F.F., See, P., Beaudin, A.E., Lum, J., Low, I., Forsberg, E.C., Poidinger, M., Zolezzi, F., Larbi, A., Ng, L.G., Chan, J.K., Greter, M., Becher, B., Samokhvalov, I.M., Merad, M., Ginhoux, F., 2015. C-Myb(+) erythro-myeloid progenitor-derived fetal monocytes give rise to adult tissue-resident macrophages. Immunity 42, 665-678.
Hoornaert, C.J., Luyckx, E., Reekmans, K., Dhainaut, M., Guglielmetti, C., Le Blon, D., Dooley, D., Fransen, E., Daans, J., Verbeeck, L., Quarta, A., De Vocht, N., Lemmens, E., Goossens, H., Van der Linden, A., Roobrouck, V.D., Verfaillie, C., Hendrix, S., Moser, M., Berneman, Z.N., Ponsaerts, P., 2016. In Vivo Interleukin-13-Primed Macrophages Contribute to Reduced Alloantigen-Specific T Cell Activation and Prolong Immunological Survival of Allogeneic Mesenchymal Stem Cell Implants. Stem Cells 34, 1971-1984.
Hu, X., Paik, P.K., Chen, J., Yarilina, A., Kockeritz, L., Lu, T.T., Woodgett, J.R., Ivashkiv, L.B., 2006. IFN-gamma suppresses IL-10 production and synergizes with TLR2 by regulating GSK3 and CREB/AP-1 proteins. Immunity 24, 563-574.
Hung, S.I., Chang, A.C., Kato, I., Chang, N.C., 2002. Transient expression of Ym1, a heparin-binding lectin, during developmental hematopoiesis and inflammation. J Leukoc Biol 72, 72-82.
Italiani, P., Boraschi, D., 2014. From Monocytes to M1/M2 Macrophages: Phenotypical vs. Functional Differentiation. Front Immunol 5, 514.
Jurewicz, M.M., Stern, L.J., 2019. Class II MHC antigen processing in immune tolerance and inflammation. Immunogenetics 71, 171-187.
Kanazawa, M., Ninomiya, I., Hatakeyama, M., Takahashi, T., Shimohata, T., 2017. Microglia and Monocytes/Macrophages Polarization Reveal Novel Therapeutic Mechanism against Stroke. Int J Mol Sci 18.
Kierdorf, K., Erny, D., Goldmann, T., Sander, V., Schulz, C., Perdiguero, E.G., Wieghofer, P., Heinrich, A., Riemke, P., Holscher, C., Muller, D.N., Luckow, B., Brocker, T., Debowski, K., Fritz, G., Opdenakker, G., Diefenbach, A., Biber, K., Heikenwalder, M., Geissmann, F., Rosenbauer, F., Prinz, M., 2013. Microglia emerge from erythromyeloid precursors via Pu.1- and Irf8-dependent pathways. Nat Neurosci 16, 273-280.
Koeniger, T., Kuerten, S., 2017. Splitting the “Unsplittable”: Dissecting Resident and Infiltrating Macrophages in Experimental Autoimmune Encephalomyelitis. Int J Mol Sci 18.
Kong, X., Gao, J., 2017. Macrophage polarization: a key event in the secondary phase of acute spinal cord injury. J Cell Mol Med 21, 941-954.
Kronenberg, G., Uhlemann, R., Richter, N., Klempin, F., Wegner, S., Staerck, L., Wolf, S., Uckert, W., Kettenmann, H., Endres, M., Gertz, K., 2018. Distinguishing features of microglia- and monocyte-derived macrophages after stroke. Acta Neuropathol 135, 551-568.
Le Blon, D., Guglielmetti, C., Hoornaert, C., Quarta, A., Daans, J., Dooley, D., Lemmens, E., Praet, J., De Vocht, N., Reekmans, K., Santermans, E., Hens, N., Goossens, H., Verhoye, M., Van der Linden, A., Berneman, Z., Hendrix, S., Ponsaerts, P., 2016. Intracerebral transplantation of interleukin 13-producing mesenchymal stem cells limits microgliosis, oligodendrocyte loss and demyelination in the cuprizone mouse model. J Neuroinflammation 13, 288.
Lively, S., Schlichter, L.C., 2018. Microglia Responses to Pro-inflammatory Stimuli (LPS, IFNgamma+TNFalpha) and Reprogramming by Resolving Cytokines (IL-4, IL-10). Front Cell Neurosci 12, 215.
Lund, H., Pieber, M., Parsa, R., Han, J., Grommisch, D., Ewing, E., Kular, L., Needhamsen, M., Espinosa, A., Nilsson, E., Overby, A.K., Butovsky, O., Jagodic, M., Zhang, X.M., Harris, R.A., 2018. Competitive repopulation of an empty microglial niche yields functionally distinct subsets of microglia-like cells. Nat Commun 9, 4845.
Luyckx, E., Van Leuven, W., Andre, D., Quarta, A., Reekmans, K., Fransen, E., Moens, L., Hankeln, T., Ponsaerts, P., Dewilde, S., 2018. Loss of Neuroglobin Expression Alters Cdkn1a/Cdk6-Expression Resulting in Increased Proliferation of Neural Stem Cells. Stem Cells Dev 27, 378-390.
Manich, G., Recasens, M., Valente, T., Almolda, B., Gonzalez, B., Castellano, B., 2019. Role of the CD200-CD200R Axis During Homeostasis and Neuroinflammation. Neuroscience 405, 118-136.
Masuch, A., van der Pijl, R., Funer, L., Wolf, Y., Eggen, B., Boddeke, E., Biber, K., 2016. Microglia replenished OHSC: A culture system to study in vivo like adult microglia. Glia 64, 1285-1297.
McQuade, A., Coburn, M., Tu, C.H., Hasselmann, J., Davtyan, H., Blurton-Jones, M., 2018. Development and validation of a simplified method to generate human microglia from pluripotent stem cells. Mol Neurodegener 13, 67.
Milich, L.M., Ryan, C.B., Lee, J.K., 2019. The origin, fate, and contribution of macrophages to spinal cord injury pathology. Acta Neuropathol 137, 785-797.
Mosser, D.M., Edwards, J.P., 2008. Exploring the full spectrum of macrophage activation. Nat Rev Immunol 8, 958-969.
Muffat, J., Li, Y., Yuan, B., Mitalipova, M., Omer, A., Corcoran, S., Bakiasi, G., Tsai, L.H., Aubourg, P., Ransohoff, R.M., Jaenisch, R., 2016. Efficient derivation of microglia-like cells from human pluripotent stem cells. Nat Med 22, 1358-1367.
Murray, P.J., 2017. Macrophage Polarization. Annu Rev Physiol 79, 541-566.
Neiva, I., Malva, J.O., Valero, J., 2014. Can we talk about microglia without neurons? A discussion of microglial cell autonomous properties in culture. Front Cell Neurosci 8, 202.
Noto, D., Sakuma, H., Takahashi, K., Saika, R., Saga, R., Yamada, M., Yamamura, T., Miyake, S., 2014. Development of a culture system to induce microglia-like cells from haematopoietic cells. Neuropathol Appl Neurobiol 40, 697-713.
Orihuela, R., McPherson, C.A., Harry, G.J., 2016. Microglial M1/M2 polarization and metabolic states. Br J Pharmacol 173, 649-665.
Pandya, H., Shen, M.J., Ichikawa, D.M., Sedlock, A.B., Choi, Y., Johnson, K.R., Kim, G., Brown, M.A., Elkahloun, A.G., Maric, D., Sweeney, C.L., Gossa, S., Malech, H.L., McGavern, D.B., Park, J.K., 2017. Differentiation of human and murine induced pluripotent stem cells to microglia-like cells. Nat Neurosci 20, 753-759.
Pasque, V., Tchieu, J., Karnik, R., Uyeda, M., Sadhu Dimashkie, A., Case, D., Papp, B., Bonora, G., Patel, S., Ho, R., Schmidt, R., McKee, R., Sado, T., Tada, T., Meissner, A., Plath, K., 2014. X chromosome reactivation dynamics reveal stages of reprogramming to pluripotency. Cell 159, 1681-1697.
Pedragosa, J., Salas-Perdomo, A., Gallizioli, M., Cugota, R., Miro-Mur, F., Brianso, F., Justicia, C., Perez-Asensio, F., Marquez-Kisinousky, L., Urra, X., Gieryng, A., Kaminska, B., Chamorro, A., Planas, A.M., 2018. CNS-border associated macrophages respond to acute ischemic stroke attracting granulocytes and promoting vascular leakage. Acta Neuropathol Commun 6, 76.
Praet, J., Santermans, E., Reekmans, K., de Vocht, N., Le Blon, D., Hoornaert, C., Daans, J., Goossens, H., Berneman, Z., Hens, N., Van der Linden, A., Ponsaerts, P., 2014. Histological characterization and quantification of cellular events following neural and fibroblast(-like) stem cell grafting in healthy and demyelinated CNS tissue. Methods Mol Biol 1213, 265-283.
Prinz, M., Priller, J., 2014. Microglia and brain macrophages in the molecular age: from origin to neuropsychiatric disease. Nat Rev Neurosci 15, 300-312.
Rajkovic, O., Potjewyd, G., Pinteaux, E., 2018. Regenerative Medicine Therapies for Targeting Neuroinflammation After Stroke. Front Neurol 9, 734.
Rath, M., Muller, I., Kropf, P., Closs, E.I., Munder, M., 2014. Metabolism via Arginase or Nitric Oxide Synthase: Two Competing Arginine Pathways in Macrophages. Front Immunol 5, 532.
Reekmans, K., De Vocht, N., Praet, J., Le Blon, D., Hoornaert, C., Daans, J., Van der Linden, A., Berneman, Z., Ponsaerts, P., 2013. Quantitative evaluation of stem cell grafting in the central nervous system of mice by in vivo bioluminescence imaging and postmortem multicolor histological analysis. Methods Mol Biol 1052, 125-141.
Reekmans, K.P., Praet, J., De Vocht, N., Tambuyzer, B.R., Bergwerf, I., Daans, J., Baekelandt, V., Vanhoutte, G., Goossens, H., Jorens, P.G., Ysebaert, D.K., Chatterjee, S., Pauwels, P., Van Marck, E., Berneman, Z.N., Van der Linden, A., Ponsaerts, P., 2011. Clinical potential of intravenous neural stem cell delivery for treatment of neuroinflammatory disease in mice? Cell Transplant 20, 851-869.
Ritzel, R.M., Patel, A.R., Grenier, J.M., Crapser, J., Verma, R., Jellison, E.R., McCullough, L.D., 2015. Functional differences between microglia and monocytes after ischemic stroke. J Neuroinflammation 12, 106.
Rock, K.L., Reits, E., Neefjes, J., 2016. Present Yourself! By MHC Class I and MHC Class II Molecules. Trends Immunol 37, 724-737.
Roszer, T., 2015. Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms. Mediators Inflamm 2015, 816460.
Saederup, N., Cardona, A.E., Croft, K., Mizutani, M., Cotleur, A.C., Tsou, C.L., Ransohoff, R.M., Charo, I.F., 2010. Selective chemokine receptor usage by central nervous system myeloid cells in CCR2-red fluorescent protein knock-in mice. PLoS One 5, e13693.
Schulz, C., Gomez Perdiguero, E., Chorro, L., Szabo-Rogers, H., Cagnard, N., Kierdorf, K., Prinz, M., Wu, B., Jacobsen, S.E., Pollard, J.W., Frampton, J., Liu, K.J., Geissmann, F., 2012. A lineage of myeloid cells independent of Myb and hematopoietic stem cells. Science 336, 86-90.
Shu, R., Bai, D., Sheu, T., He, Y., Yang, X., Xue, C., He, Y., Zhao, M., Han, X., 2017. Sclerostin Promotes Bone Remodeling in the Process of Tooth Movement. PLoS One 12, e0167312.
Smith, T.D., Tse, M.J., Read, E.L., Liu, W.F., 2016. Regulation of macrophage polarization and plasticity by complex activation signals. Integr Biol (Camb) 8, 946-955.
Stansley, B., Post, J., Hensley, K., 2012. A comparative review of cell culture systems for the study of microglial biology in Alzheimer's disease. J Neuroinflammation 9, 115.
Stoppini, L., Buchs, P.A., Muller, D., 1991. A simple method for organotypic cultures of nervous tissue. J Neurosci Methods 37, 173-182.
Takata, K., Kozaki, T., Lee, C.Z.W., Thion, M.S., Otsuka, M., Lim, S., Utami, K.H., Fidan, K., Park, D.S., Malleret, B., Chakarov, S., See, P., Low, D., Low, G., Garcia-Miralles, M., Zeng, R., Zhang, J., Goh, C.C., Gul, A., Hubert, S., Lee, B., Chen, J., Low, I., Shadan, N.B., Lum, J., Wei, T.S., Mok, E., Kawanishi, S., Kitamura, Y., Larbi, A., Poidinger, M., Renia, L., Ng, L.G., Wolf, Y., Jung, S., Onder, T., Newell, E., Huber, T., Ashihara, E., Garel, S., Pouladi, M.A., Ginhoux, F., 2017. Induced-Pluripotent-Stem-Cell-Derived Primitive Macrophages Provide a Platform for Modeling Tissue-Resident Macrophage Differentiation and Function. Immunity 47, 183-198 e186.
Timmerman, R., Burm, S.M., Bajramovic, J.J., 2018. An Overview of in vitro Methods to Study Microglia. Front Cell Neurosci 12, 242.
Varvel, N.H., Neher, J.J., Bosch, A., Wang, W., Ransohoff, R.M., Miller, R.J., Dingledine, R., 2016. Infiltrating monocytes promote brain inflammation and exacerbate neuronal damage after status epilepticus. Proc Natl Acad Sci U S A 113, E5665-5674.
Vinet, J., Weering, H.R., Heinrich, A., Kalin, R.E., Wegner, A., Brouwer, N., Heppner, F.L., Rooijen, N., Boddeke, H.W., Biber, K., 2012. Neuroprotective function for ramified microglia in hippocampal excitotoxicity. J Neuroinflammation 9, 27.
Yamasaki, R., Lu, H., Butovsky, O., Ohno, N., Rietsch, A.M., Cialic, R., Wu, P.M., Doykan, C.E., Lin, J., Cotleur, A.C., Kidd, G., Zorlu, M.M., Sun, N., Hu, W., Liu, L., Lee, J.C., Taylor, S.E., Uehlein, L., Dixon, D., Gu, J., Floruta, C.M., Zhu, M., Charo, I.F., Weiner, H.L., Ransohoff, R.M., 2014. Differential roles of microglia and monocytes in the inflamed central nervous system. J Exp Med 211, 1533-1549.
Zeger, S.L., Liang, K.Y., Albert, P.S., 1988. Models for Longitudinal Data - a Generalized Estimating Equation Approach. Biometrics 44, 1049-1060.
Zhuang, L., Pound, J.D., Willems, J.J., Taylor, A.H., Forrester, L.M., Gregory, C.D., 2012. Pure populations of murine macrophages from cultured embryonic stem cells. Application to studies of chemotaxis and apoptotic cell clearance. J Immunol Methods 385, 1-14.