Théry Manuel, Piel Matthieu
Laboratoire Biopuces, iRTSV, DSV, CEA, 38054, Grenoble, France.
Cold Spring Harb Protoc. 2009 Jul;2009(7):pdb.prot5255. doi: 10.1101/pdb.prot5255.
This protocol describes a simple, fast, and efficient method for making adhesive micropatterns that can be used to control individual cell shape and adhesion patterns. It is based on the use of an elastomeric stamp containing microfeatures to print proteins on the substrate of choice. The process can be subdivided into three parts. First, a silicon master is fabricated, which contains the microfeatures of interest. Once fabricated, the master can be used multiple times to make stamps. Masters with customized patterns can also be purchased commercially. Second, a polydimethylsiloxane (PDMS) stamp is fabricated. Unlike fabrication of the master, this step can be performed without specialized equipment. The PDMS stamp is inked with extracellular matrix proteins. Proteins are printed on a substrate (e.g., a tissue culture polystyrene dish or a glass coverslip covered with a thin layer of polystyrene). The nonprinted areas are back-filled with poly-L-lysine-polyethylene glycol, which renders them resistant to cell adhesion. The production of these micropatterned substrates can be completed in <2 h. The third and final portion of the protocol describes the deposition of cells onto the micropatterned substrate.
本方案描述了一种简单、快速且高效的制作粘性微图案的方法，该方法可用于控制单个细胞的形状和粘附模式。它基于使用含有微特征的弹性印章在所选底物上打印蛋白质。该过程可分为三个部分。首先，制作一个硅模板，其中包含感兴趣的微特征。一旦制作完成，该模板可多次用于制作印章。具有定制图案的模板也可从商业渠道购买。其次，制作聚二甲基硅氧烷（PDMS）印章。与制作模板不同，这一步无需专门设备即可完成。将细胞外基质蛋白涂覆在PDMS印章上。蛋白质被打印到底物上（例如，组织培养聚苯乙烯培养皿或覆盖有薄层聚苯乙烯的玻璃盖玻片）。未打印的区域用聚-L-赖氨酸-聚乙二醇回填，使其具有抗细胞粘附性。这些微图案化底物的制作可在2小时内完成。方案的第三部分也是最后一部分描述了将细胞沉积到微图案化底物上的过程。
