Abstract
The protocol described in this chapter covers the preparation and culture of enriched populations of microglia, astrocytes, and oligodendrocytes from the cortex and spinal cord of neonatal rat and mouse. The procedure is based on the enzymatic digestion of tissue, followed by the culture of a mixed glial cell population which is then utilized as the starting point for the isolation, via differential attachment, of the different cell types.
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Schwab ME, and Schnell L (1991) Channeling of developing rat corticospinal tract axons by myelin-associated neurite growth inhibitors. J Neurosci 11, 709–721
Rosin C, Colombo S, Calver AA, Bates TE, and Skaper SD (2005) Dopamine D2 and D3 receptor agonists limit oligodendrocyte injury caused by glutamate oxidative stress and oxygen/glucose deprivation. Glia 52, 336–343
Rosin C, Bates TE, and Skaper SD (2004) Excitatory amino acid induced oligodendrocyte cell death in vitro: receptor-dependent and-independent mechanisms. J Neurochem 90, 1173–1185
Matute C, Alberdi E, Domercq M, Perez-Cerda F, Perez-Samartin A, and Sanchez-Gomez MV (2001) The link between excitotoxic oligodendroglial death and demyelinating diseases. Trends Neurosci 24, 224–230
Gallo V, and Ghiani CA (2000) Glutamate receptors in glia: new cells, new inputs and new functions. Trends Pharmacol Sci 21, 252–258
Skaper SD, Evans NA, Soden PE, Rosin C, Facci L, and Richardson JC (2009) Oligoden-drocytes are a novel source of amyloid peptide generation. Neurochem Res 34, 2243–2250 (doi: 10.1007/s11064-009-0022-9)
Trang T, Beggs S, Wan X, and Salter MW (2009) P2X4-receptor-mediated synthesis and release of brain-derived neurotrophic factor in microglia is dependent on calcium and p38-mitogen-activated protein kinase activation. J Neurosci 29, 3518–3528
Zhang P, Hatter A, and Liu B (2007) Manganese chloride stimulates rat microglia to release hydrogen peroxide. Toxicol Lett 173, 88–100
Culbert AA, Skaper SD, Howlett DR, Evans NA, Facci L, Soden PE, et al (2006) MAPKAP kinase 2 deficiency in microglia inhibits pro-inflammatory mediator release and resultant neurotoxicity: Relevance to neuroinflammation in a transgenic mouse model of Alzheimer’s disease. J Biol Chem 281, 23658–23667
Skaper SD, Facci L, Culbert A, Evans NA, Chessell I, Davis JB, et al (2006) P2X7 receptors on microglial cells mediate injury to cortical neurons in vitro. Glia 54, 234–242
Varon S, Skaper SD, Barbin G, Selak I, and Manthorpe M (1984) Low molecular weight agents support survival of cultured neurons from the central nervous system. J Neurosci 4, 654–658
Shen LH, Li Y, and Chopp M (2010) Astrocytic endogenous glial cell derived neurotrophic factor production is enhanced by bone marrow stromal cell transplantation in the ischemic boundary zone after stroke in adult rats. Glia 58, 1074–1081
Skaper SD, Facci L, and Leon A (1995) Inflammatory mediator stimulation of astrocytes and meningeal fibroblasts induces neuronal degeneration via the nitridergic pathway. J Neurochem 64, 266–276
Miller G (2005) The dark side of glia. Science 308, 778–781
Facci L, Skaper SD, Favaron M, and Leon A (1988) A role for gangliosides in astroglial cell differentiation in vitro. J Cell Biol 106, 821–828
Gabellini N, Facci L, Milani D, Negro A, Callegaro L, Skaper S.D, et al (1991) Differences in induction of c-fos transcription by cholera toxin-derived cyclic AMP and Ca2+ signals in astrocytes and 3 T3 fibroblasts. Exp Cell Res 194, 210–217
Hamby ME, Uliasz TF, Hewett SJ, and Hewett JA (2006) Characterization of an improved procedure for the removal of microglia from confluent monolayers of primary astrocytes. J Neurosci Methods 150, 128–137
Thiele DL, Kurosaka M, and Lipsky PE (1983) Phenotype of the accessory cell necessary for mitogen-activated T and B cell responses in human peripheral blood: delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester. J Immunol 131, 2282–2290
Giulian D, Vaca K, and Corpuz M (1993) Brain glia release factors with opposing actions upon neuronal survival. J Neurosci 13, 29–37
Hewett JA, Hewett SJ, Winkler S, and Pfeiffer SE (1999) Inducible nitric oxide synthase expression in cultures enriched for mature oligodendrocytes is due to microglia. J Neurosci Res 56, 189–198
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Skaper, S.D., Argentini, C., Barbierato, M. (2012). Culture of Neonatal Rodent Microglia, Astrocytes, and Oligodendrocytes from Cortex and Spinal Cord. In: Skaper, S. (eds) Neurotrophic Factors. Methods in Molecular Biology, vol 846. Humana Press. https://doi.org/10.1007/978-1-61779-536-7_7
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DOI: https://doi.org/10.1007/978-1-61779-536-7_7
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