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Nat Protoc. 2009;4(1):78-85. doi: 10.1038/nprot.2008.199.

Primary support cultures of hippocampal and substantia nigra neurons.

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Oncology Research Unit, The Children's Hospital at Westmead, Sydney, Westmead, New South Wales 2145, Australia.


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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