Abstract
Although rodent models have been essential to unveil the emerging functions of astrocytes, the existence of interspecies differences calls for caution in extrapolating data from rodent to human astrocytes. We have developed highly enriched primary astrocyte cultures from human fetuses and adult cerebro-cortical biopsies from neurosurgery patients. Immunocytochemical characterization shows that cultures are composed of more than 95% of cells expressing in vitro astrocytic markers. Examination of the morphological and proliferative properties of cultures derived from the cerebral cortex and the hypothalamus both in untreated conditions and after treatment with EGF-related ligands illustrates the high plasticity of human astrocytes and their functional heterogeneity according to the cerebral region of origin. Our preparation offers the opportunity to characterize human astrocyte functions in vitro and also provides a valuable tool for studying the functional heterogeneity of human astrocytes isolated from distinct brain regions.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Wang, D. D., Bordey, A. (2008) The astrocyte odyssey. Prog Neurobiol 86, 342–367.
Oberheim, N. A., Takano, T., Han, X., He, W., Lin, J. H., Wang, F., Xu, Q., Wyatt, J. D., Pilcher, W., Ojemann, J. G., Ransom, B. R., Goldman, S. A., and Nedergaard, M. (2009) Uniquely hominid features of adult human astrocytes. J Neurosci 29, 3276–87.
Sharif, A., Duhem-Tonnelle, V., Allet, C., Baroncini, M., Loyens, A., Kerr-Conte, J., Collier, F., Blond, S., Ojeda, S. R., Junier, M. P., and Prevot, V. (2009) Differential erbB signaling in astrocytes from the cerebral cortex and the hypothalamus of the human brain. Glia 57, 362–79.
Zelenaia, O., Schlag, B. D., Gochenauer, G. E., Ganel, R., Song, W., Beesley, J. S., Grinspan, J. B., Rothstein, J. D., and Robinson, M. B. (2000) Epidermal growth factor receptor agonists increase expression of glutamate transporter GLT-1 in astrocytes through pathways dependent on phosphatidylinositol 3-kinase and transcription factor NF-kappaB. Mol Pharmacol 57, 667–78.
Zhou, R., Wu, X., and Skalli, O. (2001) TGF-alpha induces a stationary, radial-glia like phenotype in cultured astrocytes. Brain Res Bull 56, 37–42.
Sharif, A., Legendre, P., Prevot, V., Allet, C., Romao, L., Studler, J. M., Chneiweiss, H., and Junier, M. P. (2007) Transforming growth factor alpha promotes sequential conversion of mature astrocytes into neural progenitors and stem cells. Oncogene 26, 2695–706.
Sharif, A., Prevot, V., Renault-Mihara, F., Allet, C., Studler, J. M., Canton, B., Chneiweiss, H., and Junier, M. P. (2006) Transforming growth factor alpha acts as a gliatrophin for mouse and human astrocytes. Oncogene 25, 4076–85.
Junier, M. P. (2000) What role(s) for TGFalpha in the central nervous system? Prog Neurobiol 62, 443–73.
Rugh, R. (1964) Vertebrate embryology: the dynamics of development. Harcourt, Brace & World, New York.
Acknowledgments
We thank Pr. Serge Blond and Dr. Marc Baroncini from the Neurosurgery clinic (CHRU de Lille, Hôpital Roger Salengro, Lille, France) for giving us access to adult human cerebral cortex biopsies and Pr. Francis Collier from the gynecologic department (CHRU de Lille, Hôpital Jeanne-de-Flandre, Lille, France) for giving us access to human fetuses. We are grateful to Dr. Heintz for his generous gift of BLBP antibody. We thank Anne Loyens for expert technical help. This research was supported by Inserm Grants U816 and U837, the University of Lille 2, the Institut National du Cancer (INCa, grant PL58; Coordinator: Pierre Formstecher), the Region Nord Pas de Calais (Coordinator: Pierre Formstecher), the Agence National pour la Recherche (ANR, France to VP), the Fondation pour la Recherche Médicale (Equipe FRM, France to VP), and the Imaging Core of IFR114. Ariane Sharif was a postdoctoral fellow supported by the Région Nord Pas de Calais (2-year grant), the FRM (2-year grant), and the Fondation de France (1-year grant).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Sharif, A., Prevot, V. (2012). Isolation and Culture of Human Astrocytes. In: Milner, R. (eds) Astrocytes. Methods in Molecular Biology, vol 814. Humana Press. https://doi.org/10.1007/978-1-61779-452-0_11
Download citation
DOI: https://doi.org/10.1007/978-1-61779-452-0_11
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-451-3
Online ISBN: 978-1-61779-452-0
eBook Packages: Springer Protocols