title: Low cost production of 3D-printed devices and electrostimulation chambers for the culture of primary neurons.
abstract: The analysis of primary neurons is a basic requirement for many areas of neurobiology. However, the range of commercial systems available for culturing primary neurons is functionally limiting, and the expense of these devices is a barrier to both exploratory and large-scale studies. This is especially relevant as primary neurons often require unusual geometries and specialised coatings for optimum growth. Fortunately, the recent revolution in 3D printing offers the possibility to generate customised devices, which can support neuronal growth and constrain neurons in defined paths, thereby enabling many aspects of neuronal physiology to be studied with relative ease. In this article, we provide a detailed description of the system hardware and software required to produce affordable 3D-printed culture devices, which are also compatible with live-cell imaging. In addition, we also describe how to use these devices to grow and stimulate neurons within geometrically constrained compartments and provide examples to illustrate the practical utility and potential that these protocols offer for many aspects of experimental neurobiology.
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool, L69 3BX, United Kingdom.
authors.forename: Joanna D
authors.initials: JD
authors.lastname: Wardyn
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool, L69 3BX, United Kingdom.
authors.forename: Chris
authors.initials: C
authors.lastname: Sanderson
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool, L69 3BX, United Kingdom. Electronic address: laura.swan@liverpool.ac.uk.
authors.forename: Laura E
authors.initials: LE
authors.lastname: Swan
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Crown Street, Liverpool, L69 3BX, United Kingdom. Electronic address: massimiliano.stagi@liverpool.ac.uk.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Stagi
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D049229:Dendritic Spines / Q000648:ultrastructure; D015194:Diffusion Chambers, Culture; D004558:Electric Stimulation / Q000379:methods*; D004622:Embryo, Mammalian; D006624:Hippocampus / Q000166:cytology; D051379:Mice; D008953:Models, Anatomic; D009474:Neurons / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D066330:Printing, Three-Dimensional / Q000191:economics / Q000295:instrumentation; D014404:Tubulin / Q000378:metabolism
chemical_list: D014404:Tubulin
keywords: 3D printer; Axon; Dendrite; Live imaging; Neuronal culture; Synapse