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新型神经元培养微流控平台:神经元成分的培养和生化分析。

Novel microfluidic platform for culturing neurons: culturing and biochemical analysis of neuronal components.

机构信息

BioMEMS Team, Electronics and Telecommunication Research Institute, Daejeon, Korea.

出版信息

Biotechnol J. 2009 Nov;4(11):1573-7. doi: 10.1002/biot.200900159.

DOI:10.1002/biot.200900159
PMID:19918787
Abstract

Neurons, one of the most polarized types of cells, are typically composed of cell bodies (soma), dendrites, and axons. Many events such as electric signal transmission, axonal transport, and local protein synthesis occur in the axon, so that a method for isolating axons from somata and dendrites is required for systematically investigating these axonal events. Based on a previously developed neuron culture method for isolating and directing the growth of central nervous system axons without introducing neutrophins, we report three modified microfluidic platforms: (1) for performing biochemical analysis of the pure axonal fraction, (2) for culturing tissue explants, and (3) a design that allows high content assay on same group of cells. The key feature of these newly developed platforms is that the devices incorporate a number of microgrooves for isolating axons from the cell body. They utilize an open cellculture area, unlike the enclosed channels of the previous design. This design has extended the axonal channel so that a sufficient amount of pure axonal fraction can be obtained to perform biochemical analysis. The design also addresses the drawback of the previous neuron culture device, which was not adaptable for culturing thick neuronal tissues such as brain explants, neurospheres, and embryoid bodies, which are essential model tissues in neuroscience research. The design has an open cellculture area in the center and four enclosed channels around open area, and is suitable for multiple drug screening assays.

摘要

神经元是最具极性的细胞类型之一,通常由细胞体(胞体)、树突和轴突组成。许多事件,如电信号传递、轴突运输和局部蛋白质合成,都发生在轴突中,因此需要一种方法从胞体和树突中分离轴突,以便系统地研究这些轴突事件。基于先前开发的一种神经元培养方法,该方法可以在不引入神经营养因子的情况下分离和引导中枢神经系统轴突的生长,我们报告了三种改进的微流控平台:(1)用于对纯轴突部分进行生化分析,(2)用于培养组织外植体,以及(3)一种允许对同一组细胞进行高通量检测的设计。这些新开发的平台的关键特征是,这些设备包含了许多微沟,用于从细胞体中分离轴突。它们采用开放式细胞培养区域,与先前设计的封闭通道不同。这种设计延长了轴突通道,以便获得足够数量的纯轴突部分进行生化分析。该设计还解决了先前神经元培养设备的一个缺点,即该设备不适应培养厚的神经元组织,如脑外植体、神经球和类胚体,这些组织是神经科学研究中的重要模型组织。该设计的中心有一个开放式细胞培养区,周围有四个封闭通道,适用于多种药物筛选测定。

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