Top

Journal of Neuroinflammation

Published in:

01-12-2020 | Cytokines | Research

A primary neural cell culture model to study neuron, astrocyte, and microglia interactions in neuroinflammation

Authors: Noah Goshi, Rhianna K. Morgan, Pamela J. Lein, Erkin Seker

Published in: Journal of Neuroinflammation | Issue 1/2020

Abstract

Background

Interactions between neurons, astrocytes, and microglia critically influence neuroinflammatory responses to insult in the central nervous system. In vitro astrocyte and microglia cultures are powerful tools to study specific molecular pathways involved in neuroinflammation; however, in order to better understand the influence of cellular crosstalk on neuroinflammation, new multicellular culture models are required.

Methods

Primary cortical cells taken from neonatal rats were cultured in a serum-free “tri-culture” medium formulated to support neurons, astrocytes, and microglia, or a “co-culture” medium formulated to support only neurons and astrocytes. Caspase 3/7 activity and morphological changes were used to quantify the response of the two culture types to different neuroinflammatory stimuli mimicking sterile bacterial infection (lipopolysaccharide (LPS) exposure), mechanical injury (scratch), and seizure activity (glutamate-induced excitotoxicity). The secreted cytokine profile of control and LPS-exposed co- and tri-cultures were also compared.

Results

The tri-culture maintained a physiologically relevant representation of neurons, astrocytes, and microglia for 14 days in vitro, while the co-cultures maintained a similar population of neurons and astrocytes, but lacked microglia. The continuous presence of microglia did not negatively impact the overall health of the neurons in the tri-culture, which showed reduced caspase 3/7 activity and similar neurite outgrowth as the co-cultures, along with an increase in the microglia-secreted neurotrophic factor IGF-1 and a significantly reduced concentration of CX3CL1 in the conditioned media. LPS-exposed tri-cultures showed significant astrocyte hypertrophy, increase in caspase 3/7 activity, and the secretion of a number of pro-inflammatory cytokines (e.g., TNF, IL-1α, IL-1β, and IL-6), none of which were observed in LPS-exposed co-cultures. Following mechanical trauma, the tri-culture showed increased caspase 3/7 activity, as compared to the co-culture, along with increased astrocyte migration towards the source of injury. Finally, the microglia in the tri-culture played a significant neuroprotective role during glutamate-induced excitotoxicity, with significantly reduced neuron loss and astrocyte hypertrophy in the tri-culture.

Conclusions

The tri-culture consisting of neurons, astrocytes, and microglia more faithfully mimics in vivo neuroinflammatory responses than standard mono- and co-cultures. This tri-culture can be a useful tool to study neuroinflammation in vitro with improved accuracy in predicting in vivo neuroinflammatory phenomena.

Available only for authorised users

Corps KN, Roth TL, McGavern DB. Inflammation and neuroprotection in traumatic brain injury. JAMA Neurol. 2015;72:355–62.PubMedPubMedCentralCrossRef

Jayaraj RL, Azimullah S, Beiram R, Jalal FY, Rosenberg GA. Neuroinflammation: friend and foe for ischemic stroke. J Neuroinflammation. 2019;16:142.PubMedPubMedCentralCrossRef

Waisman A, Liblau RS, Becher B. Innate and adaptive immune responses in the CNS. Lancet Neurol [Internet]. Elsevier Ltd; 2015;14:945–55. Available from: https://doi.org/10.1016/S1474-4422(15)00141-6.

Eggen BJL, Raj D, Hanisch U. Microglial phenotype and adaptation. J Neuroimmune Pharmacol. 2013;8:807–23.PubMedCrossRef

Jensen CJ, Massie A, De Keyser J. Immune players in the CNS: the astrocyte. J Neuroimmune Pharmacol. 2013;8:824–39.PubMedCrossRef

Polikov VS, Tresco PA, Reichert WM. Response of brain tissue to chronically implanted neural electrodes. J Neurosci Methods. 2005;148:1–18.PubMedCrossRef

Miller SJ. Astrocyte heterogeneity in the adult central nervous system. Front Cell Neurosci. 2018;12:401.PubMedPubMedCentralCrossRef

Li Q, Barres BA. Microglia and macrophages in brain homeostasis and disease. Nat Rev Immunol [Internet]. Nature Publishing Group; 2017;18:225–42. Available from: https://doi.org/10.1038/nri.2017.125.

Salter MW, Stevens B. Microglia emerge as central players in brain disease. Nat Med [Internet]. Nature Publishing Group; 2017;23:1018–27. Available from: https://doi.org/10.1038/nm.4397.

10.

Jha MK, Jo M, Kim JH, Suk K. Microglia-astrocyte crosstalk: an intimate molecular conversation. Neuroscientist. 2019;25:227–40.PubMedCrossRef

11.

Szepesi Z, Manouchehrian O, Bachiller S, Deierborg T. Bidirectional microglia – neuron communication in health and disease. Front Cell Neurosci. 2018;12:323.PubMedPubMedCentralCrossRef

12.

Guttenplan KA, Liddelow SA. Astrocytes and microglia : models and tools. J Exp Med. 2018;216:71–83.PubMedCrossRef

13.

Harry GJ, Kraft AD. Neuroinflammation and microglia: considerations and approaches for neurotoxicity assessment. Expert Opin Drug Metab Toxicol. 2008;4:1265–77.PubMedPubMedCentralCrossRef

14.

Luna-Medina R, Cortes-Canteli M, Alonso M, Santos A, Perez-Castillo A. Regulation of inflammatory response in neural cells in vitro by thiadiazolidinones derivatives through peroxisome proliferator-activated receptor γ activation. J Biol Chem. 2005;280:21453–62.PubMedCrossRef

15.

Liddelow SA, Guttenplan KA, Clarke LE, Bennett FC, Bohlen CJ, Schirmer L, et al. Neurotoxic reactive astrocytes are induced by activated microglia. Nature [Internet]. Nature Publishing Group; 2017;541:481–7. Available from: http://www.nature.com/, https://doi.org/10.1038/nature21029.

16.

Culbert AA, Skaper SD, Howlett DR, Evans NA, Facci L, Soden PE, et al. MAPK-activated protein kinase 2 deficiency in microglia inhibits pro-inflammatory mediator release and resultant neurotoxicity. Relevance to neuroinflammation in a transgenic mouse model of alzheimer disease *. J Biol Chem. 2006;281:23658–67.PubMedCrossRef

17.

Gresa-Arribas N, Vieitez C, Dentesano G, Serratosa J, Saura J, Sola C. Modelling neuroinflammation in vitro: a tool to test the potential neuroprotective effect of anti-inflammatory agents. PLoS One. 2012;7.

18.

Roque PJ, Costa LG. Co-culture of neurons and microglia. Curr Protoc Toxicol. 2017;74:11–24.PubMedPubMedCentralCrossRef

19.

Bohlen CJ, Bennett FC, Tucker AF, Collins HY, Mulinyawe SB, Barres BA. Diverse requirements for microglial survival, specification, and function revealed by defined-medium cultures. Neuron [Internet]. Elsevier Inc.; 2017;94:759-773.e8. Available from: https://doi.org/10.1016/j.neuron.2017.04.043.

20.

Efremova L, Schildknecht S, Adam M, Pape R, Gutbier S, Hanf B, et al. Prevention of the degeneration of human dopaminergic neurons in an astrocyte co-culture system allowing endogenous drug metabolism. Br J Pharmacol. 2015;172:4119–32.PubMedPubMedCentralCrossRef

21.

Jones E V, Cook D, Murai KK. A Neuron-astrocyte co-culture system to investigate astrocyte-secreted factors in mouse neuronal development. Astrocytes. Humana Press; 2012. p. 341–352.

22.

Ozog MA, Siushansian R, Naus CCG. Blocked gap junctional coupling increases glutamate-induced neurotoxicity in neuron-astrocyte co-cultures. J Neuropathol Exp Neurol. 2002;61:132–41.PubMedCrossRef

23.

Mizuno T, Kuno R, Nitta A, Nabeshima T, Zhang G, Kawanokuchi J, et al. Protective effects of nicergoline against neuronal cell death induced by activated microglia and astrocytes. Brain Res. 2005;1066:78–85.PubMedCrossRef

24.

Chapman CAR, Wang L, Chen H, Garrison J, Lein PJ, Seker E. Nanoporous gold biointerfaces: modifying nanostructure to control neural cell coverage and enhance electrophysiological recording performance. Adv Funct Mater [Internet]. 2016;1604631:1604631. Available from: https://doi.org/10.1002/adfm.201604631.

25.

Chapman CAR, Chen H, Stamou M, Biener J, Biener MM, Lein PJ, et al. Nanoporous gold as a neural interface coating: effects of topography, surface chemistry, and feature size. ACS Appl Mater Interfaces. 2015;7:7093–100.PubMedPubMedCentralCrossRef

26.

Wayman GA, Bose DD, Yang D, Lesiak A, Bruun D, Impey S, et al. PCB-95 modulates the calcium-dependent signaling pathway responsible for activity-dependent dendritic growth. Environ Health Perspect. 2012;120:1003–9.PubMedPubMedCentralCrossRef

27.

Huang L-K, Wang M-JJ. Image thresholding by minimizing the measures of fuzziness. Pattern Recognit. 1995;28:41–55.CrossRef

28.

von Bartheld CS, Bahney J, Herculano-houzel S. The search for true numbers of neurons and glial cells in the human brain: a review of 150 years of cell counting. J Comp Neurol. 2016;524:3865–95.CrossRef

29.

Savchenko VL, Mckanna JA, Nikonenko IR, Skibo GG. Microglia and astrocytes in the adult rat brain: comparative immunocytochemical analysis demonstrates the efficacy of lipocortin 1 immunoreactivity. Neuroscience. 2000;96:195–203.PubMedCrossRef

30.

Nakamura Y, Si QS, Kataoka K. Lipopolysaccharide-induced microglial activation in culture: temporal profiles of morphological change and release of cytokines and nitric oxide. Neurosci Res. 1999;35:95–100.PubMedCrossRef

31.

Lehnardt S, Lachance C, Patrizi S, Lefebvre S, Follett PL, Jensen FE, et al. The toll-like receptor TLR4 is necessary for lipopolysaccharide- induced oligodendrocyte injury in the CNS. J Neurosci. 2002;22:2478–86.PubMedPubMedCentralCrossRef

32.

Batista CRA, Gomes GF, Candelario-jalil E, Fiebich BL, de Oliveira ACP. Lipopolysaccharide-induced neuroinflammation as a bridge to understand neurodegeneration. Int J Mol Sci. 2019;20:2293.PubMedCentralCrossRef

33.

Nazem A, Sankowski R, Bacher M, Al-abed Y. Rodent models of neuroinflammation for Alzheimer’s disease. J Neuroinflammation. 2015;12:74.

34.

Dutta G, Zhang P, Liu B. The lipopolysaccharide Parkinson’s disease animal model: mechanistic studies and drug discovery. Fundam Clin Pharmacol. 2008;22:453–64.PubMedPubMedCentralCrossRef

35.

Henry CJ, Huang Y, Wynne A, Hanke M, Himler J, Bailey MT, et al. Minocycline attenuates lipopolysaccharide (LPS)-induced neuroinflammation, sickness behavior, and anhedonia. J Neuroinflammation. 2008;5:1–14.CrossRef

36.

Wang X, Chen S, Ma G, Ye M, Lu G. Involvement of proinflammatory factors, apoptosis, caspase-3 activation and Ca2+ disturbance in microglia activation-mediated dopaminergic cell degeneration. Mech Ageing Dev. 2005;126:1241–54.PubMedCrossRef

37.

Nimmervoll B, White R, Yang J, An S, Henn C, Sun J, et al. LPS-induced microglial secretion of TNFα increases activity-dependent neuronal apoptosis in the neonatal cerebral cortex. Cereb Cortex. 2013;23:1742–55.PubMedCrossRef

38.

Schiweck J, Eickholt BJ, Murk K. Important shapeshifter: mechanisms allowing astrocytes to respond to the changing nervous system during development, injury and disease. Front Cell Neurosci. 2018;12:261.PubMedPubMedCentralCrossRef

39.

Liang C, Park AY, Guan J. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2007;2:329–33.PubMedCrossRef

40.

Faber-Elman A, Solomon A, Abraham JA, Marikovsky M, Schwartz M. Involvement of wound-associated factors in rat brain astrocyte migratory response to axonal injury : in vitro simulation. J Clin Invest. 1996;97:162–71.PubMedPubMedCentralCrossRef

41.

Hirano S, Yonezawa T, Hasegawa H, Hattori S, Greenhill NS, Davis PF, et al. Astrocytes express type VIII collagen during the repair process of brain cold injury. Biochem Biophys Res Commun. 2004;317:437–43.PubMedCrossRef

42.

Tecoma E, Monyer H, Goldberg MP, Choi DW. Traumatic neuronal injury in vitro is attenuated by NMDA antagonists. Neruon. 1989;2:1541–5.

43.

Boche D, Cunningham C, Gauldie J, Perry VH. Transforming growth factor-β1-mediated neuroprotection against excitotoxic injury in vivo. J Cereb Blood Flow Metabolosm. 2003;23:1174–82.CrossRef

44.

Luo J, Elwood F, Britschgi M, Villeda S, Zhang H, Ding Z, et al. Colony-stimulating factor 1 receptor (CSF1R) signaling in injured neurons facilitates protection and survival. J Exp Med. 2013;210:157–72.PubMedPubMedCentralCrossRef

45.

Mathews PM, Levy E. Cystatin C in aging and in Alzheimer’s disease. Ageing Res Rev [Internet]. Elsevier B.V.; 2016;32:38–50. Available from: https://doi.org/10.1016/j.arr.2016.06.003.

46.

Lewitt MS, Boyd GW. The role of insulin-like growth factors and insulin-like growth factor-binding proteins in the nervous system. Biochem Insights. 2019;12:1–18.

47.

Su B, Cai W, Zhang C, Martinez V, Lombet A, Perbal B. The expression of ccn3 (nov)* RNA and protein in the rat central nervous system is developmentally regulated. J Clin Pathol Mol Pathol. 2001;54:184–91.CrossRef

48.

Korecka XJA, Moloney XEB, Eggers XR, Hobo B, Scheffer S, Ras-verloop N, et al. Repulsive guidance molecule a ( RGMa ) induces neuropathological and behavioral changes that closely resemble Parkinson’s disease. J Neurosci. 2017;37:9361–79.PubMedPubMedCentralCrossRef

49.

Rosenstein JM, Krum JM, Ruhrberg C. VEGF in the nervous system. Organogenesis. 2010;6:107–14.PubMedPubMedCentralCrossRef

50.

Yang Y, Hu W, Jiang S, Wang B, Li Y, Fan C, et al. The emerging role of adiponectin in cerebrovascular and neurodegenerative diseases. Biochem Biophys Acta [Internet]. Elsevier B.V.; 2015;1852:1887–94. Available from: https://doi.org/10.1016/j.bbadis.2015.06.019.

51.

Schindowski K, Bohlen O, Von SJ, Ridder DA, Herrmann O, Schober A, et al. Regulation of GDF-15, a distant TGF-β superfamily member, in a mouse model of cerebral ischemia; 2011. p. 399–409.

52.

Goldmann T, Prinz M. Role of microglia in CNS autoimmunity. Clin Dev Immunol. 2013;2013.

53.

Ambrosini E, Columba-cabezas S, Serafini B, Muscella A, Aloisi F. Astrocytes are the major intracerebral source of macrophage inflammatory protein-3α/CCL20 in relapsing experimental autoimmune encephalomyelitis and in vitro. Glia. 2003;41:290–300.PubMedCrossRef

54.

Jha KM, Lee S, Ho D, Kook H, Park K, Lee I, et al. Diverse functional roles of lipocalin-2 in the central nervous system. Neurosci Biobehav Rev [Internet]. Elsevier Ltd; 2015;49:135–56. Available from: https://doi.org/10.1016/j.neubiorev.2014.12.006.

55.

Yu H, Liu X, Zhong Y. The effect of osteopontin on microglia. Biomed Res Int. 2017;2017.

56.

Kothur K, Wienholt L, Brilot F, Dale RC. CSF cytokines/chemokines as biomarkers in neuroinflammatory CNS disorders: a systematic review. Cytokine [Internet]. Elsevier Ltd; 2016;77:227–37. Available from: https://doi.org/10.1016/j.cyto.2015.10.001.

57.

Chitu V, Nandi S, Mehler MF, Stanley ER. Emerging roles for CSF-1 receptor and its ligands in the nervous system. Trends Neurosci. 2016;39:378–93.PubMedPubMedCentralCrossRef

58.

Elmore MRP, Najafi AR, Koike MA, Dagher NN, Spangenberg EE, Rice RA, et al. Colony-stimulating factor 1 receptor signaling is necessary for microglia viability, unmasking a microglia progenitor cell in the adult brain. Neuron [Internet]. Elsevier Inc.; 2014;82:380–97. Available from: https://doi.org/10.1016/j.neuron.2014.02.040.

59.

Erblich B, Zhu L, Etgen AM, Dobrenis K, Pollard JW. Absence of colony stimulation factor-1 receptor results in loss of microglia, Disrupted Brain Development and Olfactory Deficits. PLoS One. 2011;6:e26317.PubMedPubMedCentralCrossRef

60.

Abutbul S, Shapiro J, Szaingurten-solodkin I, Levy N, Carmy Y, Baron R, et al. TGF-β signaling through SMAD2/3 induces the quiescent microglial phenotype within the CNS environment. Glia. 2012;60:1160–71.PubMedCrossRef

61.

Butovsky O, Jedrychowski MP, Moore CS, Cialic R, Lanser AJ, Gabriely G, et al. Identification of a unique TGF-β–dependent molecular and functional signature in microglia. Nat Neurosci. 2014;17:131.PubMedCrossRef

62.

Mahley RW. Central nervous system lipoproteins: ApoE and regulation of cholesterol metabolism. Arterioscler Thromb Vasc Biol. 2016;36:1305–15.PubMedPubMedCentralCrossRef

63.

Meyers EA, Kessler JA. TGF-β family signaling in neural and neuronal differentiation, development, and function. Cold Spring Harb Perspect Biol. 2017;9.

64.

Krieglstein K, Strelau J, Schober A, Sullivan A, Unsicker K. TGF-β and the regulation of neuron survival and death. J Physiol. 2002;96:25–30.

65.

Pfrieger FW, Ungerer N. Cholesterol metabolism in neurons and astrocytes. Prog Lipid Res [Internet]. Elsevier Ltd; 2011;50:357–71. Available from: https://doi.org/10.1016/j.plipres.2011.06.002.

66.

Wang YQ, Berezovska O, Fedoroff S. Expression of colony stimulating factor-1 receptor (CSF-1R) by CNS neurons in mice. J Neurosci Res. 1999;57:616–32.PubMedCrossRef

67.

Nandi S, Gokhan S, Dai XM, Wei S, Enikolopov G, Lin H, et al. The CSF-1 receptor ligands IL-34 and CSF-1 exhibit distinct developmental brain expression patterns and regulate neural progenitor cell maintenance and maturation. Dev Biol [Internet]. Elsevier; 2012;367:100–13. Available from: https://doi.org/10.1016/j.ydbio.2012.03.026.

68.

Wei S, Nandi S, Chitu V, Yeung Y-G, Yu W, Huang M, et al. Functional overlap but differential expression of CSF-1 and IL-34 in their CSF-1 receptor-mediated regulation of myeloid cells. J Leukoc Biol. 2010;88:495–505.PubMedPubMedCentralCrossRef

69.

Lindholm D, Castrdn E, Kiefer R, Zafra F, Thoenen H. Transforming growth factor-ßl in the rat brain: increase after injury and inhibition of astrocyte proliferation. J Cell Biol. 1992;117:395–400.PubMedCrossRef

70.

Nishiyama A, Boshans L, Goncalves CM, Wegrzyn J, Patel KD. Lineage, fate, and fate potential of NG2-glia. Brain Res [Internet]. Elsevier; 2016;1638:116–28. Available from: https://doi.org/10.1016/j.brainres.2015.08.013.

71.

Wellman SM, Kozai TDY. In vivo spatiotemporal dynamics of NG2 glia activity caused by neural electrode implantation. Biomaterials [Internet]. Elsevier Ltd; 2018;164:121–33. Available from: https://doi.org/10.1016/j.biomaterials.2018.02.037.

72.

Dimou L, Gallo V. NG2-glia and their functions in the central nervous system. Glia. 2015;63:1429–51.PubMedPubMedCentralCrossRef

73.

Cuadros MA, Navascue J. The origin and differentiation of microglial cells during development. Prog Neurobiol. 1998;56:173–89.PubMedCrossRef

74.

Dalmau I, Vela JM, Gonzalez B, Finsen B, Castellano B. Dynamics of microglia in the developing rat brain. J Comp Neurol. 2003;458:144–57.PubMedCrossRef

75.

Harry GJ, Kraft AD. Microglia in the developing brain: a potential target with lifetime effects. Neurotoxicology [Internet]. Elsevier B.V.; 2012;33:191–206. Available from: https://doi.org/10.1016/j.neuro.2012.01.012.

76.

Reichert F, Rotshenker S. Galectin-3 (MAC-2) controls microglia phenotype whether amoeboid and phagocytic or branched and non-phagocytic by regulating the cytoskeleton. Front Cell Neurosci. 2019;13:90.PubMedPubMedCentralCrossRef

77.

Ueno M, Fujita Y, Tanaka T, Nakamura Y, Kikuta J, Ishii M, et al. Layer V cortical neurons require microglial support for survival during postnatal development. Nat Neurosci. Nature Publishing Group; 2013;16:543–551.

78.

Arnoux I, Audinat E. Fractalkine signaling and microglia functions in the developing brain. Neural Plast. 2015;2015.

79.

Paolicelli RC, Bisht K, Tremblay ME. Fractalkine regulation of microglial physiology and consequences on the brain and behavior. Front Cell Neurosci. 2014;8:129.PubMedPubMedCentralCrossRef

80.

Bianchi VE, Locatelli V, Rizzi L. Neurotrophic and neuroregenerative effects of GH/IGF1. Int J Mol Sci. 2017;18:2441.PubMedCentralCrossRef

81.

Papageorgiou IE, Lewen A, Galow L V., Cesetti T, Scheffel J, Regen T, et al. TLR4-activated microglia require IFN-γ to induce severe neuronal dysfunction and death in situ. Proc Natl Acad Sci [Internet]. 2016;113:212–7. Available from: https://doi.org/10.1073/pnas.1513853113.

82.

Fernández-Arjona MDM, Grondona JM, Granados-Durán P, Fernández-llebrez P, López-Ávalos MD. Microglia Morphological categorization in a rat model of neuroinflammation by hierarchical cluster and principal components analysis. Front Cell Neurosci. 2017;11:235.PubMedPubMedCentralCrossRef

83.

Abd-El-Basset E, Fedoroff S. Effect of bacterial wall lipopolysaccharide (LPS) on morphology, motility, and cytoskeletal organization of microglia in cultures. J Neurosci Res. 1995;41:222–37.PubMedCrossRef

84.

Persson M, Brantefjord M, Hansson E, Ronnback L. Lipopolysaccharide increases microglial GLT-1 expression and glutamate uptake capacity in vitro by a mechanism dependent on TNF-α. Glia. 2005;51:111–20.PubMedCrossRef

85.

Donat CK, Scott G, Gentleman SM, Sastre M. Microglial activation in traumatic brain injury. Front Aging Neurosci. 2017;9.

86.

Gulino M, Kim D, Pané S, Santos SD, Pêgo AP. Tissue response to neural implants: the use of model systems toward new design solutions of implantable microelectrodes. Front Neurosci. 2019;13:1–24.CrossRef

87.

Burda JE, Bernstein AM, Sofroniew M V. Astrocyte roles in traumatic brain injury. Exp Neurol [Internet]. Elsevier Inc.; 2016;275:305–15. Available from: https://doi.org/10.1016/j.expneurol.2015.03.020.

88.

Wang H, Song G, Chuang H, Chiu C, Abdelmaksoud A, Ye Y, et al. Portrait of glial scar in neurological diseases. Int J Immunopathol Pharmacol. 2018;31:1–6.

89.

Salatino JW, Ludwig KA, Kozai TDY, Purcell EK. Glial responses to implanted electrodes in the brain. Nat Biomed Eng. 2017;1:1.CrossRef

90.

Hu F, Ku M, Markovic D, Dildar O, Lehnardt S, Synowitz M, et al. Glioma-associated microglial MMP9 expression is upregulated by TLR2 signaling and sensitive to minocycline. Int J cancer. 2014;135:2569–78.PubMedPubMedCentralCrossRef

91.

Hsu JC, Bourguignon LYW, Adams CM, Peyrollier K, Zhang H, Fandel T, et al. Matrix metalloproteinase-9 facilitates glial scar formation in the injured spinal cord. J Neurosci. 2008;28:13467–77.PubMedPubMedCentralCrossRef

92.

Devinsky O, Vezzani A, Najjar S, Lanerolle NC De, Rogawski MA. Glia and epilepsy: excitability and inflammation. Trends Neurosci [Internet]. Elsevier Ltd; 2013;36:174–84. Available from: https://doi.org/10.1016/j.tins.2012.11.008.

93.

Schousboe A, Waagepetersen HS. Role of astrocytes in glutamate homeostasis: implications for excitotoxicity. Neurotox Res. 2005;8:221–5.PubMedCrossRef

94.

Bonde C, Sarup A, Schousboe A, Gegelashvili G, Zimmer J, Noraberg J. Neurotoxic and neuroprotective effects of the glutamate transporter inhibitor DL-threo-beta-benzyloxyaspartate (DL-TBOA) during physiological and ischemia-like conditions. Neurochem Int. 2003;43:371–80.PubMedCrossRef

95.

Samson AJ, Robertson G, Zagnoni M, Connolly CN. Neuronal networks provide rapid neuroprotection against spreading toxicity. Sci Rep [Internet]. Nature Publishing Group; 2016;6:1–11. Available from: https://doi.org/10.1038/srep33746.

96.

Eyo UB, Peng J, Murugan M, Xu P, Margolis DJ, Wu L. Regulation of physical microglia–neuron interactions by fractalkine signaling after status epilepticus. 2016;3:1–14.

97.

Vinet J, van Weering HA, Kälin RE, Wegner A, Brouwer N, et al. Neuroprotective function for ramified microglia in hippocampal excitotoxicity. J Neuroinflammation. 2012;9:27.PubMedPubMedCentralCrossRef

98.

Kato G, Inada H, Wake H, Akiyoshi R, Miyamoto A, Eto K, et al. Microglial contact prevents excess depolarization and rescues neurons from excitotoxicity. Eneuro. 2016;3:1–9.CrossRef

99.

Masuch A, Shieh C, van Rooijen N, van Calker D, Biber K. Mechanism of microglia neuroprotection: involvement of P2X 7, TNF α, and valproic acid. Glia. 2016;64:76–89.PubMedCrossRef

100.

Zhang B, Korolj A, Lai BFL, Radisic M. Advances in organ-on-a-chip engineering. Nat Rev Mater [Internet]. Springer US; 2018;3:257–78. Available from: https://doi.org/10.1038/s41578-018-0034-7.

Title: A primary neural cell culture model to study neuron, astrocyte, and microglia interactions in neuroinflammation
Authors: Noah Goshi
Rhianna K. Morgan
Pamela J. Lein
Erkin Seker
Publication date: 01-12-2020
Publisher: BioMed Central
Keyword: Cytokines
Published in: Journal of Neuroinflammation / Issue 1/2020
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-020-01819-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

A primary neural cell culture model to study neuron, astrocyte, and microglia interactions in neuroinflammation

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2020

Memantine ameliorates motor impairments and pathologies in a mouse model of neuromyelitis optica spectrum disorders

Inhibition of mast cell tryptase attenuates neuroinflammation via PAR-2/p38/NFκB pathway following asphyxial cardiac arrest in rats

LncGBP9/miR-34a axis drives macrophages toward a phenotype conducive for spinal cord injury repair via STAT1/STAT6 and SOCS3

Eicosapentaenoic acid prevents the progression of intracranial aneurysms in rats

Astragaloside IV inhibits astrocyte senescence: implication in Parkinson’s disease

DKK3 attenuates JNK and AP-1 induced inflammation via Kremen-1 and DVL-1 in mice following intracerebral hemorrhage