Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol
Author(s) -
Marco Pacifici,
Francesca Peruzzi
Publication year - 2012
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/3965
Subject(s) - nucleofection , lipofectamine , immunocytochemistry , transfection , embryonic stem cell , microbiology and biotechnology , biology , hippocampal formation , transduction (biophysics) , electrophysiology , cell culture , neuroscience , ionomycin , biochemistry , endocrinology , intracellular , gene , genetics , recombinant dna , vector (molecular biology)
We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied successfully to the isolation of mouse and human primary neurons and neural progenitors. Dissociated neurons are maintained in serum-free medium up to several weeks. These cultures can be used for nucleofection, immunocytochemistry, nucleic acids preparation, as well as electrophysiology. Older neuronal cultures can also be transfected with a good efficiency rate by lentiviral transduction and, less efficiently, with calcium phosphate or lipid-based methods such as lipofectamine.
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