Department of Chemical and Biomolecular Engineering, Yonsei University, 134 Sinchon-Dong, Seodaemoon-Gu, Seoul 120-749, Republic of Korea.
Acta Biomater. 2011 Mar;7(3):1281-9. doi: 10.1016/j.actbio.2010.11.006. Epub 2010 Nov 5.
In this paper, we describe a simple method for fabricating micropatterned nanoporous substrates that are capable of controlling the spatial positioning of mammalian cells. Micropatterned substrates were prepared by fabricating poly(ethylene glycol) (PEG) hydrogel microstructures on alumina membranes with 200 nm nanopores using photolithography. Because hydrogel precursor solution could infiltrate and become crosslinked within the nanopores, the resultant hydrogel micropatterns were firmly anchored on the substrate without the use of adhesion-promoting monolayers, thereby allow tailoring of the surface properties of unpatterned nanoporous areas. For mammalian cell patterning, arrays of microwells of different dimensions were fabricated. These microwells were composed of hydrophilic PEG hydrogel walls surrounding nanoporous bottoms that were modified with cell-adhesive Arg-Gly-Asp (RGD) peptides. Because the PEG hydrogel was non-adhesive towards proteins and cells, cells adhered selectively and remained viable within the RGD-modified nanoporous regions, thereby creating cellular micropatterns. Although the morphology of cell clusters and the number of cells inside one microwell were dependent on the lateral dimension of the microwells, adhered cells that were in direct contact with nanopores were able to penetrate into the nanopores by small extensions (filopodia) for all the different sizes of microwells evaluated.
在本文中，我们描述了一种制造能够控制哺乳动物细胞空间定位的微图案化纳米多孔基底的简单方法。通过使用光刻技术在具有 200nm 纳米孔的氧化铝膜上制造聚乙二醇（PEG）水凝胶微结构来制备微图案化基底。由于水凝胶前体溶液可以渗透并在纳米孔内交联，因此所得的水凝胶微图案在不使用促进黏附的单层的情况下牢固地固定在基底上，从而可以定制未图案化纳米多孔区域的表面性质。为了对哺乳动物细胞进行图案化，我们制造了不同尺寸的微孔阵列。这些微孔由亲水 PEG 水凝胶壁组成，水凝胶壁围绕着经过细胞黏附 Arg-Gly-Asp（RGD）肽修饰的纳米多孔底部。由于 PEG 水凝胶对蛋白质和细胞没有黏附性，因此细胞选择性地黏附并在 RGD 修饰的纳米多孔区域内保持存活，从而形成细胞微图案。尽管细胞簇的形态和一个微孔内的细胞数量取决于微孔的横向尺寸，但与纳米孔直接接触的黏附细胞能够通过小突起（纤毛）穿透所有不同尺寸的微孔。
