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Assessing Neural Stem Cell Motility Using an Agarose Gel-based Microfluidic Device

机译:使用基于琼脂糖凝胶的微流控设备评估神经干细胞运动性。

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摘要

While microfluidic technology is reaching a new level of maturity for macromolecular assays, cell-based assays are still at an infant stage1. This is largely due to the difficulty with which one can create a cell-compatible and steady microenvironment using conventional microfabrication techniques and materials. We address this problem via the introduction of a novel microfabrication material, agarose gel, as the base material for the microfluidic device. Agarose gel is highly malleable, and permeable to gas and nutrients necessary for cell survival, and thus an ideal material for cell-based assays. We have shown previously that agarose gel based devices have been successful in studying bacterial and neutrophil cell migration2. In this report, three parallel microfluidic channels are patterned in an agarose gel membrane of about 1mm thickness. Constant flows with media/buffer are maintained in the two side channels using a peristaltic pump. Cells are maintained in the center channel for observation. Since the nutrients and chemicals in the side channels are constantly diffusing from the side to center channel, the chemical environment of the center channel is easily controlled via the flow along the side channels. Using this device, we demonstrate that the movement of neural stem cells can be monitored optically with ease under various chemical conditions, and the experimental results show that the over expression of epidermal growth factor receptors (EGFR) enhances the motility of neural stem cells. Motility of neural stem cells is an important biomarker for assessing cells aggressiveness, thus tumorigenic factor3. Deciphering the mechanism underlying NSC motility will yield insight into both disorders of neural development and into brain cancer stem cell invasion.
机译:尽管微流体技术已达到大分子分析的新水平,但基于细胞的分析仍处于婴儿阶段1。这主要是由于使用传统的微细加工技术和材料很难创造出与细胞相容且稳定的微环境。我们通过引入新型微加工材料琼脂糖凝胶作为微流体装置的基础材料来解决此问题。琼脂糖凝胶具有很高的延展性,可渗透细胞存活所需的气体和营养,因此是进行基于细胞的测定的理想材料。先前我们已经表明,基于琼脂糖凝胶的装置已经成功地研究了细菌和嗜中性粒细胞的迁移2。在该报告中,在约1mm厚度的琼脂糖凝胶膜上对三个平行的微流体通道进行了构图。使用蠕动泵在两个侧面通道中保持带有介质/缓冲液的恒定流量。细胞保持在中央通道中以进行观察。由于侧通道中的营养物和化学物质不断地从侧通道扩散到中心通道,因此可以通过沿侧通道的流动来轻松控制中心通道的化学环境。使用该设备,我们证明了可以在各种化学条件下轻松地光学监测神经干细胞的运动,并且实验结果表明,表皮生长因子受体(EGFR)的过表达增强了神经干细胞的运动能力。神经干细胞的运动性是评估细胞攻击性的重要生物标志物,因此是致瘤因子3。破译NSC运动基础的机制将使人们深入了解神经发育障碍和脑癌干细胞侵袭。

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