The influence of neuronal density and maturation on network activity of hippocampal cell cultures: a methodological study.
The influence of neuronal density and maturation on network activity of hippocampal cell cultures: a methodological study.
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DOI：
10.1371/journal.pone.0083899
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Pedrocchi A
中科院分区：
文献类型：
--
作者：
Biffi E;Regalia G;Menegon A;Ferrigno G;Pedrocchi A
众所周知，细胞密度影响体外神经元网络的成熟过程。不同细胞密度培养的神经元在每个神经元上的突触数目不同，从而导致单个神经元的突触传递不同，从而导致密度依赖的神经元网络活动。尽管许多作者提供了关于细胞密度对神经元培养活性的影响的详细信息，但关于密度和年龄对神经元培养活性的影响的专门报道尚未见报道。因此，本工作旨在为开展海马神经元培养实验研究的研究人员提供参考数据，为实验的规划和解码提供帮助。在这项工作中，我们分析了神经元密度和培养年龄对成熟海马区培养的功能属性的影响。我们研究了三种不同细胞密度下培养的神经元的电生理活性，并用微电极阵列(MEA)记录了它们在成熟后的自发电活动。我们收集了86个独立的海马区培养的数据，以获得可靠的统计结果，考虑到文化之间的高度变异性。根据简单的尖峰、突发和网络突发特征来评估网络活动。我们观察到，电描述子的特征是在成熟过程中出现一个功能峰值，然后是一个稳定阶段(对于稀疏和中等密度培养)或一个下降阶段(对于高密度神经元培养)。此外，900个/mm2的培养显示出适合于长期实验(例如药物治疗的慢性效应)的特性，而1800个/mm2的培养应该更适合于需要强烈电活动的实验(例如评估抑制分子的效果)。最后，3600个/mm2的细胞培养更适合于与节省时间相关的实验(例如药物筛选)。这些结果旨在为MEAs的体外神经生理学和神经药理学实验的规划提供参考。
It is known that cell density influences the maturation process of in vitro neuronal networks. Neuronal cultures plated with different cell densities differ in number of synapses per neuron and thus in single neuron synaptic transmission, which results in a density-dependent neuronal network activity. Although many authors provided detailed information about the effects of cell density on neuronal culture activity, a dedicated report of density and age influence on neuronal hippocampal culture activity has not yet been reported. Therefore, this work aims at providing reference data to researchers that set up an experimental study on hippocampal neuronal cultures, helping in planning and decoding the experiments. In this work, we analysed the effects of both neuronal density and culture age on functional attributes of maturing hippocampal cultures. We characterized the electrophysiological activity of neuronal cultures seeded at three different cell densities, recording their spontaneous electrical activity over maturation by means of MicroElectrode Arrays (MEAs). We had gather data from 86 independent hippocampal cultures to achieve solid statistic results, considering the high culture-to-culture variability. Network activity was evaluated in terms of simple spiking, burst and network burst features. We observed that electrical descriptors were characterized by a functional peak during maturation, followed by a stable phase (for sparse and medium density cultures) or by a decrease phase (for high dense neuronal cultures). Moreover, 900 cells/mm2 cultures showed characteristics suitable for long lasting experiments (e.g. chronic effect of drug treatments) while 1800 cells/mm2 cultures should be preferred for experiments that require intense electrical activity (e.g. to evaluate the effect of inhibitory molecules). Finally, cell cultures at 3600 cells/mm2 are more appropriate for experiments in which time saving is relevant (e.g. drug screenings). These results are intended to be a reference for the planning of in vitro neurophysiological and neuropharmacological experiments with MEAs.
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