title: Microfluidics and multielectrode array-compatible organotypic slice culture method.
abstract: Organotypic brain slice cultures are used for a variety of molecular, electrophysiological, and imaging studies. However, the existing culture methods are difficult or expensive to apply in studies requiring long-term recordings with multielectrode arrays (MEAs). In this work, a novel method to maintain organotypic cultures of rodent hippocampus for several weeks on standard MEAs in an unmodified tissue culture incubator is described. Polydimethylsiloxane (Sylgard) mini-wells were used to stabilize organotypic cultures on glass and MEA surfaces. Hippocampus slices were successfully maintained within PDMS mini-wells for multiple weeks, with preserved pyramidal layer organization, connectivity, and activity. MEAs were used to record the development of spontaneous activity in an organotypic cultures for 4 weeks. This method is compatible with integration of microchannels into the culture substrate. Microchannels were incorporated into the mini-wells and applied to the guidance of axons originating within the slice, paving the way for studies of axonal sprouting using organotypic slices.
authors.affiliation: Center for Engineering in Medicine, Department of Surgery, Harvard Medical School, Massachusetts General Hospital, Boston, MA, United States.
authors.forename: Yevgeny
authors.initials: Y
authors.lastname: Berdichevsky
authors.forename: Helen
authors.initials: H
authors.lastname: Sabolek
authors.forename: John B
authors.initials: JB
authors.lastname: Levine
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Staley
authors.forename: Martin L
authors.initials: ML
authors.lastname: Yarmush
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D004558:Electric Stimulation / Q000379:methods; D005452:Fluoresceins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000502:physiology; D008839:Microelectrodes; D044085:Microfluidics / Q000379:methods*; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000502:physiology; D009924:Organ Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D005452:Fluoresceins; C007740:fluorexon