A device for co-culturing autonomic neurons and cardiomyocytes using micro-fabrication techniques

Abstract

It is still unclear how the activity of sympathetic and parasympathetic neurons influences the activity of cardiomyocytes in culture. We developed a device for co-culturing sympathetic neurons, parasympathetic neurons, and cardiomyocytes using micro-fabrication techniques. Morphological connections between each type of autonomic neuron and the cardiomyocytes were observed by immunostaining. The inter-beat-interval (IBI) of the cardiomyocytes was modulated after electrically stimulating each type of autonomic neuron. Modulation of the IBI was blocked by the addition of pharmacological blockers to the culture medium. These results suggest that the co-culture device can be utilized to understand how the activity of sympathetic neurons and parasympathetic neurons influences the activity of cardiomyocytes in the culture environment.

Conditioned medium of sympathetic ganglia with addition of fibroblast growth factor 2 promotes plasticity of cardiomyocytes in culture

“…Such findings are corroborated by several studies that point to a precise and intrinsic relationship between the sympathetic system and the heart, in which there is a "neuro-cardiac" junction, where sympathetic neurotransmission would occur in a pathway similar to neuronal synapses (Conforti, Tohse & Sperelakis, 1991;Zaglia & Mongillo, 2017), besides the presence of postganglionic neurons projecting to various areas of the heart (Kucera & Hrabovska, 2015). In addition, the effects of sympathetic and parasympathetic neurons play an important role in cardiomyocyte culture (Oiwa et al, 2016).…”

Instrumented Microphysiological Systems for Real-Time Measurement and Manipulation of Cellular Electrochemical Processes

“…utilized a >11,000 MEA to study signal propagation along constrained rat cortical neurites in PDMS microchannels (Lewandowska et al., 2015). This platform allowed for the recording of discrete action potentials propagating from the soma to individual neurites or the axonal terminus.

Figure 3Examples of Instrumented Organ-on-chip Models of the Peripheral and Central Nervous System(A and B) Extracellular activity and connectivity of compartmentalized cortical neurons via an MEA.(C and D) Extracellular recordings of neural activity and 3D connectivity via a 3D MEA.(E) Extracellular recordings and impedance detection to quantify firing frequency and cell adhesion of neuronal cells exposed to sodium valproic acid via an MEA and an interdigitated electrode structure.(F) Extracellular recordings of spontaneous and stimulated autonomic neuron activity for regulating cardiac beating via an MEA.(G) CV of myelinated and nonmyelinated sensory neurons via a microchannel-electrode approach.(H) Single-unit action potentials and CV of intact sciatic nerves exposed to ultrasound stimuli via a multi-wire electrode array.(I) CAP and CV of intact sciatic nerves via a microchannel-electrode approach.(J, K, and L) (J) CAP recording in 3D rodent sensory neuron, (K) 3D myelinated rodent sensory neurons, and (L) 3D human stem cell-derived sensory neuron models via stimulating and recording wire electrodes.Reprinted and adapted with permission from: A (Kanagasabapathi et al., 2011); B (Pan et al., 2015); C (Rowe et al., 2007); D (Musick et al., 2009); E (Koester et al., 2010a); F (Oiwa et al., 2016); G (Sakai et al., 2017); H (Chen et al., 2017); I (Gribi et al., 2018); J (Huval et al., 2015); K (Khoshakhlagh et al., 2018); and L (Sharma et al., 2019). MEA, multielectrode array; CV, conduction velocity; CAP, compound action potential.

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“…Reprinted and adapted with permission from: A (Kanagasabapathi et al., 2011); B (Pan et al., 2015); C (Rowe et al., 2007); D (Musick et al., 2009); E (Koester et al., 2010a); F (Oiwa et al., 2016); G (Sakai et al., 2017); H (Chen et al., 2017); I (Gribi et al., 2018); J (Huval et al., 2015); K (Khoshakhlagh et al., 2018); and L (Sharma et al., 2019). MEA, multielectrode array; CV, conduction velocity; CAP, compound action potential.…”

“…Pulsatile electrical stimulation (Oiwa et al., 2016), pharmacological stimulation (Smith et al., 2013), and light stimulation of transgenic neurons (Uzel et al., 2016) have been used to evoke neural firing and confirm functional synapsing onto muscle cells. The displacement of passive force transducers (Morimoto et al., 2013, Smith et al., 2013, Uzel et al., 2016), relative fluorescent intensity of Ca 2+ gradients (Ionescu et al., 2016, Zahavi et al., 2015), and myocyte electrophysiology (Oiwa et al., 2016, Takeuchi et al., 2011) have all been used as quantitative measures of muscle contraction frequency and/or strength in response to spontaneous and/or evoke neurogenic control. Further, in addition to contractile properties, myoblasts function as endocrine organs and release cytokines following physical exertion (Pedersen and Febbraio, 2008).…”

“…Oiwa et al. established an MPS model of the CNS in which both neural and cardiac electrophysiology was recorded via an integrated MEA (Figure 3F) (Oiwa et al., 2016). Cardiac and neural populations were physically constrained into separate compartments for independent assessment.…”