Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale.
Exploiting the superior protein resistance of polymer brushes to control single cell adhesion and polarisation at the micron scale.
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DOI：
10.1016/j.biomaterials.2010.02.066
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发表时间：
2010-06
期刊：
影响因子：
14
通讯作者：
Huck, Wilhelm T. S.
中科院分区：
文献类型：
--
作者：
Gautrot, Julien E.;Trappmann, Britta;Oceguera-Yanez, Fabian;Connelly, John;He, Ximin;Watt, Fiona M.;Huck, Wilhelm T. S.
在模型图案化底物上对细胞微环境的控制允许在良好定义的条件下系统地研究细胞生物学，可能使用自动化系统。利用聚低聚（甲基丙烯酸乙二醇酯）（POEGMA）刷的极端蛋白质抗性来实现单细胞的高保真模式。这些涂层可以通过软光刻在大面积（显微镜载玻片）和规模（基片通常以200批为单位制备）上进行图案化。目前的方案依赖于细胞外基质（ECM）蛋白在无保护区域的吸附，使用简单的孵育和洗涤步骤。通过椭偏仪和SPR检测，发现POEGMA刷在储存和细胞培养过程中都具有优异的稳定性。通过荧光显微镜和原子力显微镜研究了基材处理、电刷厚度和孵育方案对超薄金基材和玻璃基材ECM沉积的影响。优化的条件导致高质量的微米级ECM模式，甚至在玻璃基板上，适合于控制细胞扩散和极化。这些模式与用于活细胞成像的最先进技术（荧光显微镜，FRET）兼容。该技术与单细胞分析方法相结合，为探索调节细胞行为的机制提供了一个平台。
The control of the cell microenvironment on model patterned substrates allows the systematic study of cell biology in well defined conditions, potentially using automated systems. The extreme protein resistance of poly(oligo(ethylene glycol methacrylate)) (POEGMA) brushes is exploited to achieve high fidelity patterning of single cells. These coatings can be patterned by soft lithography on large areas (a microscope slide) and scale (substrates were typically prepared in batches of 200). The present protocol relies on the adsorption of extra-cellular matrix (ECM) proteins on unprotected areas using simple incubation and washing steps. The stability of POEGMA brushes, as examined via ellipsometry and SPR, is found to be excellent, both during storage and cell culture. The impact of substrate treatment, brush thickness and incubation protocol on ECM deposition, both for ultra-thin gold and glass substrates, is investigated via fluorescence microscopy and AFM. Optimised conditions result in high quality ECM patterns at the micron scale, even on glass substrates, that are suitable for controlling cell spreading and polarisation. These patterns are compatible with state-of-the-art technologies (fluorescence microscopy, FRET) used for live cell imaging. This technology, combined with single cell analysis methods, provides a platform for exploring the mechanisms that regulate cell behaviour.
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DOI：
10.1083/jcb.200812034
发表时间：
2009-06-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Dupin I;Camand E;Etienne-Manneville S
通讯作者：
Etienne-Manneville S
DOI：
10.1126/science.1171643
发表时间：
2009-06-26
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Discher DE;Mooney DJ;Zandstra PW
通讯作者：
Zandstra PW
影响因子：
3.9
作者：
Ma, HW;Li, DJ;Chilkoti, A
通讯作者：
Chilkoti, A
影响因子：
9.5
作者：
Gautrot, Julien E.;Huck, Wilhelm T. S.;Ramstedt, Madeleine
通讯作者：
Ramstedt, Madeleine
影响因子：
3.9
作者：
Jeon, NL;Finnie, K;Nuzzo, RG
通讯作者：
Nuzzo, RG