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用于研究趋化性过程中细胞迁移的开放式微流控装置。

Open access microfluidic device for the study of cell migration during chemotaxis.

机构信息

Department of Biological Sciences, Vanderbilt University, VU Station B #351634, Nashville, TN 37235, USA.

出版信息

Integr Biol (Camb). 2010 Nov;2(11-12):648-58. doi: 10.1039/c0ib00110d. Epub 2010 Oct 15.

DOI:10.1039/c0ib00110d
PMID:20949221
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3806978/
Abstract

Cells sense and interpret chemical gradients, and respond by localized responses that lead to directed migration. An open microfluidic device (OMD) was developed to provide quantitative information on both the gradient and morphological changes that occurred as cells crawled through various microfabricated channels. This device overcame problems that many current devices have been plagued with, such as complicated cell loading, media evaporation and channel blockage by air bubbles. We used a micropipette to set up stable gradients formed by passive diffusion and thus avoided confounding cellular responses produced by shear forces. Two versions of the OMD are reported here: one device that has channels with widths of 6, 8, 10 and 12 μm, while the other has two large 100 μm channels to minimize cellular interaction with lateral walls. These experiments compared the migration rates and qualitative behavior of Dictyostelium discoideum cells responding to measurable cAMP and folic acid gradients in small and large channels. We report on the influence that polarity has on a cell's ability to migrate when confined in a channel. Polarized cells that migrated to cAMP were significantly faster than the unpolarized cells that crawled toward folic acid. Unpolarized cells in wide channels often strayed off course, yet migrated faster than unpolarized cells in confined channels. Cells in channels farthest from the micropipette migrated through the channels at rates similar to cells in channels with higher concentrations, suggesting that cell speed was independent of mean concentration. Lastly, it was found that the polarized cells could easily change migration direction even when only the leading edge of the cell was exposed to a lateral gradient.

摘要

细胞感知和解释化学梯度,并通过局部反应做出响应,从而导致定向迁移。开发了开放式微流控装置 (OMD),以提供有关细胞在通过各种微加工通道爬行时发生的梯度和形态变化的定量信息。该装置克服了许多当前设备存在的问题,例如复杂的细胞加载、培养基蒸发和气泡堵塞通道。我们使用微吸管建立了由被动扩散形成的稳定梯度,从而避免了由剪切力产生的混杂细胞反应。这里报告了两种版本的 OMD:一种设备具有 6、8、10 和 12 μm 宽的通道,另一种设备具有两个大的 100 μm 通道,以最大程度减少细胞与侧壁的相互作用。这些实验比较了响应小通道和大通道中可测量的 cAMP 和叶酸梯度的 Dictyostelium discoideum 细胞的迁移率和定性行为。我们报告了极性对细胞在通道中受限时迁移能力的影响。向 cAMP 迁移的极化细胞比向叶酸爬行的非极化细胞快得多。宽通道中的非极化细胞经常偏离方向,但比受限通道中的非极化细胞迁移得更快。离微吸管最远的通道中的细胞以与通道中浓度较高的细胞相似的速率穿过通道,这表明细胞速度与平均浓度无关。最后,发现即使只有细胞的前缘暴露于侧向梯度,极化细胞也可以轻松改变迁移方向。

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