Abstract
We describe here a method to obtain primary cell cultures from the cerebral cortex and the hypothalamus of bovine fetuses. We report how tissue origin, developmental stages, and culture medium conditions influence cell differentiation and the prevalence of glial cells vs. neurons. We compare explants from early, middle, and late stages of development and two different fetal calf serum concentrations (1 and 10%) to identify the best conditions to obtain and grow viable astrocytes in culture. In addition, we describe how to cryopreserve and obtain viable cortical astrocytes from frozen fetal bovine brain samples.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Fournier JG, Escaig-Haye F, Grigoriev V (2000) Ultrastructural localization of prion proteins: physiological and pathological implications. Microsc Res Tech 50:76–88
Liberski PP, Brown P (2004) Astrocytes in transmissible spongiform encephalopathies (prion diseases). Folia Neuropathol 42 (Suppl B):71–88
Collinge J. (2001) Prions diseases of humans and animals: Their causes and molecular basis. Annu Rev Neurosci 24:519–550
Peruffo A, Massimino ML, Ballarin C, Carmignoto G, Rota A, Cozzi B (2004) Primary cultures from fetal bovine brain. Neuroreport 15:1719–22.
Peruffo A, Buson G, Cozzi B, Ballarin C. (2008) Primary cell cultures from fetal bovine hypothalamus and cerebral cortex: a reliable model to study P450Arom and alpha and beta estrogen receptors in vitro. Neurosci Lett 434:83–7.
McGeady TA, Quinn PJ, Patrick ESF and Ryan MT (2006) Veterinary Embryology. 1st edn.Blackwell Publishing Ltd., UK.
Cozzi B (1986) Cell types in the pineal gland of the horse: an ultrastructural and immunocytochemical study. Anat Rec 216: 165–174.
Eng LF, Vanderhaeghen JJ, Bignami A, Gerstl B (1971) An acidic protein isolated from fibrous astrocytes. Brain Res 28:351–4.
Acknowledgments
The authors wish to thank Dr. Livio Corain of the Department of Statistics of the University of Padua, for help with statistical analysis of data. This work was supported by the Italian Ministry for University and Research (Grants No 2002073381002 and No 60A08-3811/08).
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
Ballarin, C., Peruffo, A. (2012). Primary Cultures of Astrocytes from Fetal Bovine Brain. In: Milner, R. (eds) Astrocytes. Methods in Molecular Biology, vol 814. Humana Press. https://doi.org/10.1007/978-1-61779-452-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-61779-452-0_9
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-451-3
Online ISBN: 978-1-61779-452-0
eBook Packages: Springer Protocols