Microfluidic multicompartment device for neuroscience research
Microfluidic multicompartment device for neuroscience research
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DOI：
10.1021/la026417v
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发表时间：
2003-03-04
期刊：
影响因子：
3.9
通讯作者：
Jeon, NL
中科院分区：
文献类型：
--
作者：
Taylor, AM;Rhee, SW;Jeon, NL
本文描述和表征了一种新型的微制造神经元培养装置。该装置结合了微加工，微流体和表面微图案化技术，以创建多隔室神经元培养装置，可用于许多神经科学研究应用。该设备是在聚（二甲基硅氧烷），PDMS，使用软光刻技术。将PDMS装置置于组织培养皿（聚苯乙烯）或玻璃基底上，形成两个隔室，每个隔室的体积小于2穆尔。这两个隔室由物理屏障分开，其中嵌入了许多微米尺寸的凹槽，以允许神经突穿过隔室生长，同时保持流体隔离。将细胞接种到体（细胞体）隔室中，3-4天后，神经突通过凹槽延伸到神经炎隔室中。在培养7天后，装置中神经元的活力在50%至70%之间;这略低于但与在组织培养皿上生长的对照值相当。健康的神经元形态在器械和对照品中均很明显。我们证明了使用静水压力的能力，以隔离侮辱一个隔间，因此，暴露局部地区的神经元的侮辱适用于可溶性形式。由于微槽对液体运输的高阻力，损伤被包含在神经炎隔室中，而没有明显的渗漏进入体隔室超过15小时。最后，我们展示了使用多聚赖氨酸图案化与微加工设备相结合，以促进识别和可视化的神经元。通过微图案化技术指导神经元附着位点和神经突生长方向的能力，与培养区域内的流体隔离隔室相结合，提供了优于标准开放培养方法和用于操纵不同神经元微环境的其他常规方法的显著优势。
This paper describes and characterizes a novel microfabricated neuronal culture device. This device combines microfabrication, microfluidic, and surface micropatterning techniques to create a multicompartment neuronal culturing device that can be used in a number of neuroscience research applications. The device is fabricated in poly(dimethylsiloxane), PDMS, using soft lithography techniques. The PDMS device is placed on a tissue culture dish (polystyrene) or glass substrate, forming two compartments with volumes of less than 2 muL each. These two compartments are separated by a physical barrier in which a number of micron-size grooves are embedded to allow growth of neurites across the compartments while maintaining fluidic isolation. Cells are plated into the somal (cell body) compartment, and after 3-4 days, neurites extend into the neuritic compartment via the grooves. Viability of the neurons in the devices is between 50 and 70% after 7 days in culture; this is slightly lower than but comparable to values for a control grown on tissue culture dishes. Healthy neuron morphology is evident in both the devices and controls. We demonstrate the ability to use hydrostatic pressure to isolate insults to one compartment and, thus, expose localized areas of neurons to insults applied in soluble form. Due to the high resistance of the microgrooves for fluid transport, insults are contained in the neuritic compartment without appreciable leakage into the somal compartment for over 15 h. Finally, we demonstrate the use of polylysine patterning in combination with the microfabricated device to facilitate identification and visualization of neurons. The ability to direct sites of neuronal attachment and orientation of neurite outgrowth by micropatterning techniques, combined with fluidically isolated compartments within the culture area, offers significant advantages over standard open culture methods and other conventional methods for manipulating distinct neuronal microenvironments.