Peer-Reviewed Journal Details
Mandatory Fields
Kaar, Aidan; Morley, Shane J.; Rae, Mark G.
2017
May
Journal of Neuroscience Methods
An Efficient and Cost-Effective Method of Generating Post-Natal (P2-14) Mouse Primary Hippocampal Neuronal Cultures.
In Press
WOS: 1 ()
Optional Fields
Primary neuronal culture Hippocampal neurons Postnatal Trituration Mouse
286
69
77
Background: Primary culture of postnatal central neurons is a widely used methodology for applications such as the investigation of neuronal development, protein trafficking/distribution and cellular signalling. However, successful production and maintenance of such cultures, particularly from postnatal animals, can be challenging. In attempting to surmount these difficulties, several disparate culturing methodologies have been developed. Such methodologies are centred on the identification and optimisation of critical steps and, as such, the protocols and reagents utilised can differ quite markedly from protocol to protocol, often with the suggestion that the use of a (usually expensive) proprietary reagent(s), lengthy substrate preparation and/or cell isolation techniques is/are necessary for successful culture preparation. New method: Herein, we present a simple and inexpensive protocol for the preparation of primary hippocampal neurons from postnatal (25 day old) mice, which remain viable for experimental use for over one month. Results: Neurons cultured using this method follow well established developmental norms and display typical responses to standard physiological stimuli such as depolarisation and certain pharmacological agents. Comparison with existing methods/conclusion: By using a novel trituration technique, simplified methodology and non-proprietary reagents, we have developed a reliable protocol that enables the cost effective and efficient production of high quality postnatal mouse hippocampal cultures. This method, if required, can also be utilised to prepare neurons both from other regions of the brain as well as from other species such as rat.
0165-0270
10.1016/j.jneumeth.2017.05.020
Grant Details
The Physiological Society
Seed Fund Grant