Abstract
Primary cultures of rat and murine hippocampal neurons are widely used to reveal cellular mechanisms in neurobiology. Their use is limited, as culturing at low density is often not possible or is dependent on sophisticated methods. Here we present a novel method for culturing embryonic (E16.5) murine hippocampal neurons, using a spatially separated ring of cortical neurons for neurotrophic support. This method allows long-term cultures at a very low cell density, and therefore, the study of single embryo preparations and isolated neurons. This method has been adopted for neurons from the substantia nigra (E16.5), with support from a ring of striatal neurons.
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Acknowledgements
We thank Mian Bi, Fabien Delerue and Denise Nergenau for technical assistance and Dr Victor Anggono for the dynamin antibody. L.M.I. has been supported by the DFG, the Australian Research Council (ARC) and the University of Sydney. J.G. is a Medical Foundation Fellow and has been supported by the University of Sydney, the National Health and Medical Research Council (NHMRC), the ARC, the New South Wales Government through the Ministry for Science and Medical Research (BioFirst Program), the Nerve Research Foundation, the Medical Foundation (University of Sydney) and the Judith Jane Mason and Harold Stannett Williams Memorial Foundation. P.G. is supported by a NSW cancer institute research infrastructure grant and is a Principal Research Fellow of the NHMRC.
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Fath, T., Ke, Y., Gunning, P. et al. Primary support cultures of hippocampal and substantia nigra neurons. Nat Protoc 4, 78–85 (2009). https://doi.org/10.1038/nprot.2008.199
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DOI: https://doi.org/10.1038/nprot.2008.199
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