Advertisement

Aggregating Neural Cell Cultures

  • Paul Honegger
  • Florianne Monnet-Tschudi
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Aggregating brain cell cultures are primary, three-dimensional cell cultures consisting of even-sized, spherical structures that are maintained in suspension by constant gyratory agitation. Because of the avidity of freshly dissociated embryonic cells to attach to their counterparts, cell aggregates form spontaneously and rapidly under appropriate culture conditions. The reaggregated cells are able to migrate within the formed structures, and to interact with each other by direct cell-cell contact, as well as through exchange of nutritional and signaling factors. This tissue-specific environment enables aggregating neural cells to differentiate, and to develop specialized structures (e.g., synapses, myelinated axons) resembling those of brain tissue in situ. Aggregating cell cultures are therefore classified as organotypic cultures (Doyle et al., 1994).

Keywords

Cholera Toxin Cryostat Section Cytosine Arabinoside Gelatin Capsule Laminar Flow Hood 
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. Adams, J. (1981), Heavy metal intensification of DAB-based HRP reaction product. J. Histochem. Cytochem. 29, 775.PubMedGoogle Scholar
  2. Appel, K., Honegger, P., and Gebicke-Haerter, P. (1995), Expression of interleukin-3 and tumor necrosis factor-b mRNA in cultured microglia. J. Neuroimmunol. 60, 83–91.PubMedCrossRefGoogle Scholar
  3. Ashwell, K. (1990), The distribution of microglia and cell death in the developing mouse cerebellum. Dev. Brain Res. 55, 219–230.CrossRefGoogle Scholar
  4. Bardoscia, M. T., Amstad, P., and Honegger, P. (1992), Expression of the proto-oncogene c-fos in three-dimensional fetal brain cell cultures and the lack of correlation with maturation-inducing stimuli. Mol. Brain Res. 12, 23–30.PubMedCrossRefGoogle Scholar
  5. Bignami, A. and Aschner, R. (1992), Some observations on the localization of hyaluronic acid in adult, newborn and embryonal rat brain. Int. J. Dev. Neurosci. 10, 45–57.PubMedCrossRefGoogle Scholar
  6. Bradford, M. M. (1976), A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein dye binding. Analyt. Biochem. 72, 248–254.PubMedCrossRefGoogle Scholar
  7. Choi, H. K., Won, L., and Heller, A. (1993), Dopaminergic neurons grown in three-dimensional reaggregate culture for periods of up to one year. J. Neurosci. Methods 46, 233–244.PubMedCrossRefGoogle Scholar
  8. Chomczynski, P. and Sacchi, N. (1987), Single-step method of RNA isolation by acid guanidinium thiocyanatephenol-chloroform extraction. Analyt. Biochem. 162, 156–159.PubMedCrossRefGoogle Scholar
  9. Corthésy-Theulaz, I., Merillat, A.-M., Honegger, P., and Rossier, B. C. (1990), Na+-K+-ATPase gene expression during in vitro development of rat fetal forebrain. Am. J. Physiol. 258 (Cell Physiol. 27), C1062–C1069.PubMedGoogle Scholar
  10. DeLong, G. R. and Sidman, R. L. (1970), Alignment defect of reaggregating cells in cultures of developing brains of reeler mutant mice. Dev. Biol. 22, 584–599.CrossRefGoogle Scholar
  11. Downs, T. R. and Wilfinger, W. W. (1983), Fluorimetric quantification of DNA in cells and tissues. Analyt. Biochem. 131, 538–547.PubMedCrossRefGoogle Scholar
  12. Doyle, A., Griffiths, J. B., and Newell, D. G. (Principal Editors) (1994), Cell and Tissue Culture: Laboratory Procedures. Wiley, Chichester.Google Scholar
  13. Guentert-Lauber, B., Monnet-Tschudi, E., Omlin, F. X., Favrod, P., and Honegger, P. (1985), Serum-free aggregate cultures of rat CNS glial cells: biochemical, immunocytochemical and morphological characterization. Dev. Neurosci. 7, 33–44.PubMedCrossRefGoogle Scholar
  14. Honegger, P., Lenoir, D., and Favrod, P. (1979), Growth and differentiation of aggregating fetal brain cells in a serum-free defined medium. Nature 282, 305–308.PubMedCrossRefGoogle Scholar
  15. Honegger, P. and Pardo, B. (1999), Separate neuronal and glial Na+,K+-ATPase isoforms regulate glucose utilization in response to membrane depolarization and elevated extracellular potassium. J. Cerebr. Blood FlowMetabol. 19, 1051–1059.CrossRefGoogle Scholar
  16. Honegger, P. and Richelson, E. (1976), Biochemical differentiation of mechanically dissociated mammalian brain in aggregating cell culture. Brain Res. 109, 335–354.PubMedCrossRefGoogle Scholar
  17. Honegger, P. and Richelson E. (1977), Biochemical differentiation of aggregating cell cultures of different fetal rat brain regions. Brain Res. 133, 329–339.PubMedCrossRefGoogle Scholar
  18. Honegger, P and Richelson, E. (1979), Neurotransmitter synthesis, storage and release by aggregating cell cultures of rat brain. Brain Res. 162, 89–101.PubMedCrossRefGoogle Scholar
  19. Honegger, P. and Schilter, B. (1992), Serum-free aggregate cultures of fetal rat brain and liver cells: methodology and some practical applications in neurotoxicology, in: The Brain in Bits and Pieces. In vitro Techniques in Neurobiology, Neuropharmacology and Neurotoxicology, Zbinden, G., ed., MTC Verlag, Zollikon, Switzerland, pp. 51–79.Google Scholar
  20. Honegger, P. and Werffeli, P. (1988), Use of aggregating cell cultures for toxicological studies. Experientia 44, 817–823.PubMedCrossRefGoogle Scholar
  21. Huber, G. and Matus, A. (1984), Differences in the cellular distribution of two microtubule-associated proteins, MAPI and MAP2, in rat brain. J. Neurosci. 4, 151–160.PubMedGoogle Scholar
  22. Juurlink, B. H., Schousboe, A., Jorgensen, O. S., and Hertz, L. (1981), Induction by hydrocortisone of glutamine synthetase in mouse primary astrocytes. J. Neurochem. 36, 136–142.PubMedCrossRefGoogle Scholar
  23. Kerlero de Rosbo, N., Honegger, P., Lassmann, H., and Matthieu, J.-M. (1990), Demyelination induced in aggregating brain cell cultures by a monoclonal antibody against myelin/oligodendrocyte glycoprotein. J. Neurochem. 55, 583–587.PubMedCrossRefGoogle Scholar
  24. Lennette, D. A. (1978), An improved mounting medium for immunofluorescence microscopy. Am. J. Clin. Pathol. 69, 647,648.Google Scholar
  25. Lenoir, D. and Honegger, P. (1983), Insulin-like growth factor I (IGF I) stimulates DNA synthesis in fetal rat brain cell cultures. Dev. Brain Res. 7, 205–213.CrossRefGoogle Scholar
  26. Linington, C., Webb, M., and Woodhams, P. L. (1984), A novel myelin-associated glycoprotein defined by a mouse monoclonal antibody. J. Neuroimmunol. 6, 387–396.CrossRefGoogle Scholar
  27. Matthieu, J.-M., Honegger, P., Favrod, P., Gautier, E., and Dolivo, M. (1979), Biochemical characterization of a myelin fraction isolated from rat brain aggregating cell cultures. J. Neurochem. 32, 869–881.PubMedCrossRefGoogle Scholar
  28. Monnet-Tschudi, F., Zurich, M.-G., Pithon, E., van Melle, G., and Honegger, P. (1995), Microglial responsiveness as a sensitive marker for trimethyltin (TMT) neurotoxicity. Brain Res. 690, 8–14.PubMedCrossRefGoogle Scholar
  29. Moscona, A. A. (1961), Rotation-mediated histogenetic aggregation of dissociated cells: a quantifiable approach to cell interactions in vitro. Exp. Cell Res. 22, 455–475.PubMedCrossRefGoogle Scholar
  30. Pan, L. C. and Price, P. A. (1984), The effect of transcriptional inhibitors on the bone gamma-carboxyglutamic acid protein response to 1,25-dihydroxyvitamin D3 in osteosarcoma cells. J. Biol. Chem. 259, 5844–5847.PubMedGoogle Scholar
  31. Riederer, B., Cohen, R., and Matus, A. (1986), MAP5: a novel microtubule-associated protein under strong developmental regulation. J. Neurocytol. 15, 763–775.PubMedCrossRefGoogle Scholar
  32. Riederer, B. M., Monnet-Tschudi, F., and Honegger, P. (1992), Development and maintenance of the neuronal cytoskeleton in aggregated cell cultures of fetal rat telencephalon and influence of elevated K+ concentrations. J. Neurochem. 58, 649–658.PubMedCrossRefGoogle Scholar
  33. Riederer, B. M., Porchet, R., Marugg, R. A., and Binder, L. I. (1993), Solubility of cytoskeletal protein in immunohistochemistry and the influence of fixation. J. Histochem. Cytochem. 41, 609–616.PubMedGoogle Scholar
  34. Rose, S. P. R. (1965), Preparation of enriched fractions from cerebral cortex containing isolated, metabolically active neuronal cells. Nature (London) 206, 621,622.Google Scholar
  35. Schrier, B. K. (1973), Surface culture of fetal mammalian brain cells: effect of subculture on morphology and choline acetyltransferase. J. Neurobiol. 4, 117–124.PubMedCrossRefGoogle Scholar
  36. Seeds, N. W. (1971), Biochemical differentiation in reaggregating brain cell culture. Proc. Natl. Acad. Sci. USA 68, 1858–1861.PubMedCrossRefGoogle Scholar
  37. Streit, W. J. and Kreutzberg, G. W. (1987), Lectin binding by resting and reactive microglia. J. Neurocytol. 16, 249–260.PubMedCrossRefGoogle Scholar
  38. Taylor, V, Miescher, G. C., Pfarr, S., Honegger, P., Breitschopf, H., Lassmann, H., and Steck, A. J. (1994), Expression and developmental regulation of Ehk-1, a neuronal Elk-like receptor tyrosine kinase in brain. Neuroscience 63, 163–178.PubMedCrossRefGoogle Scholar
  39. Varon, S. and Raiborn, C. W., Jr. (1969), Dissociation, fractionation, and culture of embryonic brain cells. Brain Res. 12, 180–199.PubMedCrossRefGoogle Scholar
  40. Wilson, S. H., Schrier, B. K., Farber, J.-L., Thompson, E. J., Rosenberg, R. N., Blume A. J., and Nierenberg, M. W. (1972), Markers for gene expression in cultured cells from the nervous system. J. Biol. Chem. 247, 3159–3169.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Paul Honegger
    • 1
  • Florianne Monnet-Tschudi
    • 1
  1. 1.Institut de Physiologie, Faculté de MédecineUniversite de LausanneLausanneSwitzerland

Personalised recommendations