title: Preparation of neuronal co-cultures with single cell precision.
abstract: Microfluidic embodiments of the Campenot chamber have attracted great interest from the neuroscience community. These interconnected co-culture platforms can be used to investigate a variety of questions, spanning developmental and functional neurobiology to infection and disease propagation. However, conventional systems require significant cellular inputs (many thousands per compartment), inadequate for studying low abundance cells, such as primary dopaminergic substantia nigra, spiral ganglia, and Drosophilia melanogaster neurons, and impractical for high throughput experimentation. The dense cultures are also highly locally entangled, with few outgrowths (<10%) interconnecting the two cultures. In this paper straightforward microfluidic and patterning protocols are described which address these challenges: (i) a microfluidic single neuron arraying method, and (ii) a water masking method for plasma patterning biomaterial coatings to register neurons and promote outgrowth between compartments. Minimalistic neuronal co-cultures were prepared with high-level (>85%) intercompartment connectivity and can be used for high throughput neurobiology experiments with single cell precision.
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS.
authors.forename: Ngoc-Duy
authors.initials: ND
authors.lastname: Dinh
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS; Department of Biochemical Engineering, University College London.
authors.forename: Ya-Yu
authors.initials: YY
authors.lastname: Chiang
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS.
authors.forename: Heike
authors.initials: H
authors.lastname: Hardelauf
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS.
authors.forename: Sarah
authors.initials: S
authors.lastname: Waide
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS.
authors.forename: Dirk
authors.initials: D
authors.lastname: Janasek
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften, ISAS; Institute for Life Sciences, University of Southampton; J.J.West@soton.ac.uk.
authors.forename: Jonathan
authors.initials: J
authors.lastname: West
mesh_terms: D002460:Cell Line; D018920:Coculture Techniques / Q000295:instrumentation* / Q000379:methods*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D009474:Neurons / Q000166:cytology*
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon