Shrirao Anil B, Kung Frank H, Yip Derek, Firestein Bonnie L, Cho Cheul H, Townes-Anderson Ellen
Department of Biomedical Engineering, Rutgers University;
Department of Cell Biology and Neuroscience, Rutgers University;
J Vis Exp. 2017 Feb 26(120):55513. doi: 10.3791/55513.
Substrate and cell patterning techniques are widely used in cell biology to study cell-to-cell and cell-to-substrate interactions. Conventional patterning techniques work well only with simple shapes, small areas and selected bio-materials. This article describes a method to distribute cell suspensions as well as substrate solutions into complex, long, closed (dead-end) polydimethylsiloxane (PDMS) microchannels using negative pressure. This method enables researchers to pattern multiple substrates including fibronectin, collagen, antibodies (Sal-1), poly-D-lysine (PDL), and laminin. Patterning of substrates allows one to indirectly pattern a variety of cells. We have tested C2C12 myoblasts, the PC12 neuronal cell line, embryonic rat cortical neurons, and amphibian retinal neurons. In addition, we demonstrate that this technique can directly pattern fibroblasts in microfluidic channels via brief application of a low vacuum on cell suspensions. The low vacuum does not significantly decrease cell viability as shown by cell viability assays. Modifications are discussed for application of the method to different cell and substrate types. This technique allows researchers to pattern cells and proteins in specific patterns without the need for exotic materials or equipment and can be done in any laboratory with a vacuum.
底物和细胞图案化技术在细胞生物学中被广泛用于研究细胞间以及细胞与底物间的相互作用。传统的图案化技术仅适用于简单形状、小面积区域以及特定的生物材料。本文介绍了一种利用负压将细胞悬液以及底物溶液分配到复杂、长且封闭（死端）的聚二甲基硅氧烷（PDMS）微通道中的方法。该方法使研究人员能够对多种底物进行图案化，包括纤连蛋白、胶原蛋白、抗体（Sal-1）、聚-D-赖氨酸（PDL）和层粘连蛋白。底物的图案化使得人们能够间接对多种细胞进行图案化。我们已经测试了C2C12成肌细胞、PC12神经细胞系、胚胎大鼠皮质神经元以及两栖类视网膜神经元。此外，我们证明了该技术可以通过对细胞悬液短暂施加低真空在微流控通道中直接对成纤维细胞进行图案化。如细胞活力测定所示，低真空不会显著降低细胞活力。文中讨论了将该方法应用于不同细胞和底物类型时的改进措施。这项技术使研究人员能够以特定图案对细胞和蛋白质进行图案化，无需使用特殊材料或设备，并且在任何配备有真空设备的实验室中都可以完成。
