Development of Semi-Separated Co-Culture System for Electrical Stimulation and Extracellular Recording of Sympathetic Neuron and Cardiomyocyte

Abstract

A semi-separated co-culture system for spatio-temporal recording of the electrical activities from superior cervical ganglion (SCG) neurons and ventricular myocytes (VMs) was developed by using a handmade “H-shaped” chamber placed on microelectrode arrays (MEA). The chamber was made of polydimethylsyloxane (PDMS) and consisted of two chambers which were connected with a pathway. 16-20 hours after dissemination of SCG neurons into one chamber, the dissociated VMs were disseminated into the other chamber. 4days after dissemination of VMs, SCG neurons and VMs conjugated only at the pathway. Spontaneous electrical activities of SCG neurons and VMs were observed several days after the dissemination of VMs. Constant-voltage stimualtion (1 V, 1 ms, biphasic square pulses) was applied to SCG neurons at the frequency of 10 Hz using 32 electrodes. After applying electrical stimulation to SCG neurons, the contraction rate of VMs in three samples increased by 55±5.6%, 64±8.8%, 280±160%, respectively. This result suggests that neuromuscular junctions were formed between SCG neurons and VMs.