title: Device for co-culture of sympathetic neurons and cardiomyocytes using microfabrication.
abstract: Rat superior cervical ganglion (SCG) neurons and ventricular myocytes (VMs) were co-cultured separately in a minichamber placed on a microelectrode-array (MEA) substrate. The minichamber, fabricated photolithographically using polydimethylsiloxane (PDMS), had 2 compartments, 16 microcompartments and 8 microconduits. The SCG neurons were seeded into one of the compartments and all of the microcompartments using a glass pipette controlled by a micromanipulator and a microinjector. The VMs were seeded into the other compartment. Three days after seeding of the VMs, the SCG neurons were still confined to one compartment and all of the microcompartments, and the neurites of the SCG neurons had connected with the VMs via the microconduits. Constant-voltage stimulation, using a train of biphasic square pulses (1 ms at +1 V, followed by -1 ms at 1 V), was applied to the SCG neurons in the microcompartments using 16 electrodes. Evoked responses were observed in several electrodes while electrical stimulation was applied to the SCG neurons. Two-way analysis of variance (ANOVA) revealed that the frequency of the stimulation pulses had significant effects in increasing the beat rate of the VMs, and that the interaction between the frequency and the number of the pulses also had a significant effect on the ratio. No significant increases in the beat rate were observed when propranolol, a β-adrenergic receptor antagonist, was added to the culture medium. These results suggest that synaptic pathways were formed between the SCG neurons and the VMs, and that this co-culture device can be utilized for studies of network-level interactions between sympathetic neurons and cardiomyocytes.
authors.affiliation: Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba 277-8563, Japan. takeuchi@bmpe.k.u-tokyo.ac.jp
authors.forename: Akimasa
authors.initials: A
authors.lastname: Takeuchi
authors.forename: Shingo
authors.initials: S
authors.lastname: Nakafutami
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Tani
authors.forename: Masahide
authors.initials: M
authors.lastname: Mori
authors.forename: Yuzo
authors.initials: Y
authors.lastname: Takayama
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Moriguchi
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Kotani
authors.forename: Keiko
authors.initials: K
authors.lastname: Miwa
authors.forename: Jong-kook
authors.initials: JK
authors.lastname: Lee
authors.forename: Makoto
authors.initials: M
authors.lastname: Noshiro
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Jimbo
mesh_terms: D000818:Animals; D018920:Coculture Techniques / Q000295:instrumentation*; D004558:Electric Stimulation; D006352:Heart Ventricles / Q000166:cytology; D055616:Microtechnology / Q000295:instrumentation*; D032383:Myocytes, Cardiac / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D051381:Rats; D017208:Rats, Wistar; D017783:Superior Cervical Ganglion / Q000166:cytology*