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논문 기본 정보

자료유형
학술저널
저자정보
Vernekar Varadraj N. (Georgia Institute of Technology) LaPlaca Michelle C. (Georgia Institute of Technology)
저널정보
대한의용생체공학회 Biomedical Engineering Letters (BMEL) Biomedical Engineering Letters (BMEL) Vol.10 No.4
발행연도
2020.1
수록면
579 - 591 (13page)

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Three-dimensional (3-D) neural cultures represent a promising platform for studying disease and drug screening. Tools and methodologies for measuring the electrophysiological function in these cultures are needed. Therefore, the purpose of this work was primarily to develop a methodology to interface engineered 3-D dissociated neural cultures with commercially available 3-D multi-electrode arrays (MEAs) reliably over 3 weeks to enable the recording of their electrophysiological activity. We further compared the functional output of these cultures to their structural and synaptic network development over time. We reliably interfaced a primary rodent neuron-astrocyte (2:1) 3-D co-culture (2500 cells/mm3 plating cell density) in Matrigel™ (7.5 mg/mL) that was up to 750 µm thick (30–40 cell-layers) with spiked 3-D MEAs while maintaining high viability. Using these MEAs we successfully recorded the spontaneous development of neural network-level electrophysiological activity and measured the development of putative synapses and neuronal maturation in these co-cultures using immunocytochemistry over 3 weeks in vitro. Planar (2-D) MEAs interfaced with these cultures served as recording controls. Neurons within this interfaced 3-D culture-MEA system exhibited considerable neurite outgrowth, networking, neuronal maturation, synaptogenesis, and culture-wide spontaneous firing of synchronized spikes and bursts of action potentials. Network-wide spikes and synchronized bursts increased rapidly (first detected at 2 days) during the first week in culture, plateaued during the second week, and reduced slightly in the third week, while maintaining high viability throughout the 3-week culturing period. Early electrophysiology activity occurred prior to neuronal process maturation and significant synaptic density increases in the second week. We successfully interfaced 3-D neural co-cultures with 3-D MEAs and recorded the electrophysiological activity of these cultures over 3 weeks. The initial period of rapid increase in electrophysiological activity, followed by a period of neuronal maturation and high-level of synapse formation in these cultures suggests a developmental homeostatic process. This methodology can enable future applications both in fundamental investigations of neural network behavior and in translational studies involving drug testing and neural interfacing.
#Three-dimensional Electrophysiology Multi-electrode array MEA In vitro Neuron Astrocyte Synapse Matrigel Culture

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참고문헌 (72)

참고문헌 신청
Novak JL / 1988 / Multisite hippocampal slice recording and stimulation using a 32 element microelectrode array / J Neurosci Methods 23(2):149~159 google schola De Simoni A / 2003 / Development of rat CA1 neurones in acute versus organotypic slices : role of experience in synaptic morphology and activity / J Physiol 550(1):135~147 google schola Beggs JM / 2004 / Neuronal avalanches are diverse and precise activity patterns that are stable for many hours in cortical slice cultures / J Neurosci 24(22):5216~5229 google schola Wagenaar D / 2006 / An extremely rich repertoire of bursting patterns during the development of cortical cultures / BMC Neurosci 7(1):1~18 google schola Gähwiler B / 1997 / Organotypic slice cultures : a technique has come of age / Trends Neurosci 20(10):471~477 google schola

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