Issue 13, 2011

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.

Graphical abstract: Device for co-culture of sympathetic neurons and cardiomyocytes using microfabrication

Article information

Article type
Paper
Submitted
24 Aug 2010
Accepted
12 Apr 2011
First published
13 May 2011

Lab Chip, 2011,11, 2268-2275

Device for co-culture of sympathetic neurons and cardiomyocytes using microfabrication

A. Takeuchi, S. Nakafutami, H. Tani, M. Mori, Y. Takayama, H. Moriguchi, K. Kotani, K. Miwa, J. Lee, M. Noshiro and Y. Jimbo, Lab Chip, 2011, 11, 2268 DOI: 10.1039/C0LC00327A

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