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Cultures of Astroglia and Microglia from Primary Cultures of Mouse Neopallium

  • Sergey Fedoroff
  • Arleen Richardson
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Brains of newborn rats or mice are generally used as the source of tissue for glial cultures. Only about 1% of cells survive the cell disaggregation process and culture environment, and neurons that survive die within the first few days of culturing. Such cultures contain progenitor cells and glia cells in different stages of differentiation (Alliot et al.,1991; Fedoroff et al., 1997; see also  Chapter 14). Terminally differentiated glia cells, which can no longer divide, are overgrown by the proliferating, immature cells. How the cultures will develop, and which cell types (astroglia, oligodendroglia, ependymal cells, or microglia) will predominate, depend on the culture medium and the physical conditions under which the cells are grown (see also  Chapters 12 and  14). The medium can be modified either by changing its chemically defined components or by adding or deleting serum. It is also possible to add growth factors and/or cytokines, either to the culture medium in pure recombinant form, or as medium conditioned by cells that produce and secrete growth factors/cytokines into the medium (the latter is considerably cheaper). The addition of cytokines to cultures can have a dramatic effect on the morphology and function of cells. It should be noted that cytokines and growth factors may affect more than one cell type, and may initiate variable effects in different cell types. The growth factors/cytokines may interact with other factors in the medium synergistically, additively, or in an inhibitory way. It is important to remember that the half-life of cytokines is short. However, the cytokines/growth factors can be added to ,the culture in microcapsules, which release the factors at a constant rate over a long period of time, thus assuring a constant concentration of a given factor to culture medium over long periods of time (Maysinger et al., 1992; Maysinger et al., 1996a, Maysinger et al., 1996b).

Keywords

Olfactory Bulb Cerebral Hemisphere Ependymal Cell Pasteur Pipet Newborn Mouse 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Further Reading

  1. Abd-El-Basset, E. and Fedoroff, S. (1995), Effect of bacterial wall lipopolysaccharide (LPS) on morphology, motility and cytoskeletal organization of microglia in cultures. J. Neurosci. Res. 41, 222–237.PubMedCrossRefGoogle Scholar
  2. Abd-El-Basset, E. and Fedoroff, S. (1994), Dynamics of actin filaments in microglia during Fc receptor-mediated phagocytosis. Acta Neuropathol. 88, 527–537.PubMedCrossRefGoogle Scholar
  3. Alliot, F., Lecain, E., Grima, B., and Pessac, B. (1991), Microglial progenitors with a high proliferative potential in the embryonic and adult mouse brain. Proc. Natl. Acad. Set, USA, 88, 1541–1545.CrossRefGoogle Scholar
  4. Branch, R. D. and Guilbert, L. J. (1986), Practical in vitro assay systems for the measurement of hematopoietic growth factors. J. Tissue Culture Methods 10, 101–108.CrossRefGoogle Scholar
  5. Das, S. K., Stanley, E. R., Guilbert, L. J., and Farman, L. W. (1980), Determination of a colony stimulating factor subclass by a specific receptor on a macrophage cell line. J. Cell Physiol. 104, 359–366.PubMedCrossRefGoogle Scholar
  6. Fedoroff, S. (1995), Development of microglia, in Neuroglia, Kettenmann, H. and Ransom, B. R., eds., Oxford University Press, New York, pp. 162–181.Google Scholar
  7. Fedoroff, S., Hao, C., Ahmed, J., and Guilbert, L. J. (1993), Paracrine and autocrine signaling in regulation of microglia survival, in: Biology and Pathology of Astrocyte-Neuron Interactions, Fedoroff, S., Juurlink, B. H. J., and Doucette, R., eds. Plenum, New York, pp. 247–261.Google Scholar
  8. Fedoroff, S., Zhai, R., and Novak, J. P. (1997), Microglia and astroglia have a common progenitor cell. J. Neurosci. Res. 50, 477–486.PubMedCrossRefGoogle Scholar
  9. Gebicke-Haerter, P. J., Baker, J., Schobert, A., and Northoff, H. (1989), Lipopolysaccharide-free conditions in primary astrocyte cultures allow growth and isolation of microglial cells. J. Neurosci. Res. 9, 183–194.Google Scholar
  10. Giulian, D. and Bauer, T. J. (1986), Characterization of ameboid microglia isolated from developing mammalian brain. J. Neurosci. 6, 2163–2178.PubMedGoogle Scholar
  11. Hao, C., Richardson, A., and Fedoroff, S. (1991), Macrophage-like cells originate from neuroepithelium in culture: characterization and properties of the macrophage-like cells. Int. J. Dev. Neurosci. 9, 1–14.PubMedCrossRefGoogle Scholar
  12. Hao, C., Guilbert, L. J., and Fedoroff, S. (1990), Production of colony-stimulating factor-1 (CSF-1) by mouse astroglia in vitro. J. Neurosci. Res. 27, 314–323.PubMedCrossRefGoogle Scholar
  13. Hayes, G. M., Woodroofe, M. N., and Cuzner, M. L. (1988), Characterization of microglia isolated from adult human and rat brain. J. Neuroimmunol. 19, 177–189.PubMedCrossRefGoogle Scholar
  14. Maysinger, D., Jalsenjak, I., and Cuello, C. (1992), Microencapsulated nerve growth factor: effects on the forebrain neurons following devascularizing cortical lesions. Neuroscience Lett. 140, 71–74.CrossRefGoogle Scholar
  15. Maysinger, D., Berezovskaya, O., and Fedoroff, S. (1996a), The hematopoietic cytokine colony stimulating factor 1 is also growth factor in the CNS: µI) Microencapsulated CSF-1 and LM-10 cells as delivery systems. Exp. Neurol. 141, 47–56.PubMedCrossRefGoogle Scholar
  16. Maysinger, D., Krieglstein, K., Filipovic-Grcic, J., Sendtner, M., Unsicher, K., and Richardson, P. (1996b), Microencapsulated ciliary neurotrophic factor physical properties and biological activities. Exp. Neurol. 138, 177–188.PubMedCrossRefGoogle Scholar
  17. Neuhaus, J. and Fedoroff, S. (1994), Development of microglia in mouse neopallial cell cultures. Glia 11, 11–17.PubMedCrossRefGoogle Scholar
  18. Northoff, H., Gluck, D. Wolpl, A., Kubanek, B., and Galanos, C. (1986a), Lipopolysaccharide induced elaboration of interleukin-1 µL-1) by human monocytes: Use for the detection of LPS in serum and influence of serum-LPS interactions. Rev. Infect. Dis. 9(Suppl. 5), 599–602.Google Scholar
  19. Northoff, H., Kabelits, D., and Galanos, C. (1986b), Interleukin 1 production for detection of bacterial polysaccharide in fetal calf sera and other solutions. Immunol. Today 7, 126–127.CrossRefGoogle Scholar
  20. Novak, J. P. and Fedoroff, S. (1999), Model of the dynamics of a branching system of the glial cell lineages in vitro. J. Biol. Systems 7, 429–447.CrossRefGoogle Scholar
  21. Rieske, E., Graeber, M. B., Tetzlaff, W., Czlonkowska, A., Streit, W. J., and Kreutzberg, G. W. (1989), Microglia and microglia-derived brain macrophages in culture: generation from axotomized rat facial nuclei, identification and characterization in vitro. Brain Res. 429, 1–14.CrossRefGoogle Scholar
  22. Richardson, A., Hao, C., and Fedoroff, S. (1993), Microglia progenitor cells: a subpopulation in cultures of mouse neopallial astroglia. Glia 7, 25–33.PubMedCrossRefGoogle Scholar
  23. Sultzer, B. M., Castagna, R., Bandekar, J., and Wong, P. (1993), Lipopolysaccharide nonresponder cells: The C3H/HeJ defect. Immunobiol. 187, 257–271.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Sergey Fedoroff
    • 1
  • Arleen Richardson
    • 1
  1. 1.Department of Anatomy and Cell Biology, College of MedicineUniversity of SaskatchewanSaskatoonCanada

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