Microfluidics and multielectrode array-compatible organotypic slice culture method.
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.
Berdichevsky
Yevgeny
Y
Center for Engineering in Medicine, Department of Surgery, Harvard Medical School, Massachusetts General Hospital, Boston, MA, United States.
Sabolek
Helen
H
Levine
John B
JB
Staley
Kevin J
KJ
Yarmush
Martin L
ML
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
D005452:Fluoresceins; C007740:fluorexon