Institute for Bioengineering of Catalonia, C/Baldiri Reixac 11-15, 08028 Barcelona, Spain.
Langmuir. 2012 Sep 25;28(38):13688-97. doi: 10.1021/la3025638. Epub 2012 Sep 12.
In this work, we propose an easy method to produce highly tunable gradients of covalently bound proteins on topographically modified poly(methyl methacrylate). We used a microfluidic approach to obtain linear gradients with high slope (0.5 pmol·cm(-2)·mm(-1)), relevant at the single-cell level. These protein gradients were characterized using fluorescence microscopy and surface plasmon resonance. Both experimental results and theoretical modeling on the protein gradients generated have proved them to be highly reproducible, stable up to 7 days, and easily tunable. This method enables formation of versatile cell culture platforms combining both complex biochemical and physical cues in an attempt to approach in vitro cell culture methods to in vivo cellular microenvironments.
在这项工作中，我们提出了一种在经过形貌修饰的聚甲基丙烯酸甲酯上制备共价结合蛋白的高度可调梯度的简单方法。我们使用微流控方法获得了具有高斜率（0.5 pmol·cm(-2)·mm(-1)）的线性梯度，这在单细胞水平上是相关的。使用荧光显微镜和表面等离子体共振对这些蛋白梯度进行了表征。蛋白梯度的实验结果和理论模型都证明了它们具有高度的重现性，在 7 天内保持稳定，并且易于调节。该方法可形成多功能细胞培养平台，将复杂的生化和物理线索结合在一起，试图使体外细胞培养方法接近体内细胞微环境。
