Uniform cell seeding and generation of overlapping gradient profiles in a multiplexed microchamber device with normally-closed valves
Uniform cell seeding and generation of overlapping gradient profiles in a multiplexed microchamber device with normally-closed valves
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DOI：
10.1039/c0lc00086h
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Mosadegh, Bobak;Agarwal, Mayank;Takayama, Shuichi
在微流控装置中产生稳定的可溶性因子梯度可以研究各种细胞事件，如趋化和分化。然而，许多梯度装置直接将细胞暴露在持续的流体流动中，这可能会由于剪切力而引起细胞的不良反应和/或洗出细胞分泌的分子。虽然已经有了无流动梯度的设备，但它们通常只产生单一条件和/或具有不适合长期实验的衰减梯度分布。在这里，我们描述了一种微设备，它在无流动微室的单个平台上产生几个化学梯度条件，从而促进稳态梯度分布。该设备包含嵌入式常闭阀门，可以同时快速、均匀地将细胞接种到所有微腔。微通道网络将所需的解决方案从易于进入的开放式储罐分配到单个输出端口，从而实现了在设备中诱导流动的简单设置。嵌入的多孔过滤器，夹在微通道网络和细胞微室之间，允许生物分子的扩散，但阻止细胞上的任何体积流动。
Generation of stable soluble-factor gradients in microfluidic devices enables studies of various cellular events such as chemotaxis and differentiation. However, many gradient devices directly expose cells to constant fluid flow and that can induce undesired responses from cells due to shear stress and/or wash out of cell-secreted molecules. Although there have been devices with flow-free gradients, they typically generate only a single condition and/or have a decaying gradient profile that does not accommodate long-term experiments. Here we describe a microdevice that generates several chemical gradient conditions on a single platform in flow-free microchambers which facilitates steady-state gradient profiles. The device contains embedded normally-closed valves that enable fast and uniform seeding of cells to all microchambers simultaneously. A network of microchannels distributes desired solutions from easy-access open reservoirs to a single output port, enabling a simple setup for inducing flow in the device. Embedded porous filters, sandwiched between the microchannel networks and cell microchambers, enable diffusion of biomolecules but inhibit any bulk flow over the cells.