title: Generation of microisland cultures using microcontact printing to pattern protein substrates.
abstract: The capacity to isolate small numbers of neurons in vitro is an essential tool to study the cell biology of synapses and the development of neuronal networks by specific cell types. Microisland culture assays allow for single neurons, or simple neural networks, to be isolated on islands of glial cells; however, the techniques commonly used to produce microisland substrates are expensive, challenging to control, and typically result in many discarded substrates. Here, we used microcontact printing to pattern a glass surface with islands of extracellular matrix proteins known to support neural cell growth and differentiation. To promote segregation of the cells to the islands, the substrate surrounding the islands was backfilled with polyethylene glycol (PEG), forming a relatively non-permissive surface on which cell attachment is limited. Astrocytes, and subsequently hippocampal neurons, were then seeded onto the islands of patterned protein. Using this method, readily reproducible patterns of protein islands were produced that permit cell attachment, differentiation, and growth. The technique is a rapid, inexpensive, and reliable means to generate patterned substrates appropriate for microisland cultures.
authors.affiliation: McGill Program in NeuroEngineering, Department of Neurology and Neurosurgery, Montreal Neurological Institute, McGill University, 3801 University Ave., Montreal, Quebec H3A 2B4, Canada.
authors.forename: Sébastien G
authors.initials: SG
authors.lastname: Ricoult
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Goldman
authors.forename: David
authors.initials: D
authors.lastname: Stellwagen
authors.forename: David
authors.initials: D
authors.lastname: Juncker
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Kennedy
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000502:physiology*; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible / Q000737:chemistry / Q000378:metabolism; D018920:Coculture Techniques / Q000379:methods*; D016326:Extracellular Matrix Proteins / Q000737:chemistry* / Q000378:metabolism*; D054802:Molecular Imprinting / Q000379:methods*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D051381:Rats; D013499:Surface Properties
chemical_list: D020099:Coated Materials, Biocompatible; D016326:Extracellular Matrix Proteins