Jang Kihoon, Sato Kae, Mawatari Kazuma, Konno Tomohiro, Ishihara Kazuhiko, Kitamori Takehiko
Department of Applied Chemistry, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan.
Biomaterials. 2009 Mar;30(7):1413-20. doi: 10.1016/j.biomaterials.2008.11.013. Epub 2008 Dec 10.
This report describes a new surface-treatment technique for cell micropatterning. Cell attachment was selectively controlled on the glass surface using a photochemical reaction. This strategy is based on combining 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which is known to reduce non-specific adsorption, and a photolabile linker (PL) for selective cell patterning. The MPC polymer was coated directly on the glass surface using a straightforward surface modification method, and was removed by ultraviolet (UV) light illumination. All the surface modification steps were evaluated using static water contact angle measurements, X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), measurements of non-specific protein adsorption, and the cell attachment test. After selective cleavage of the MPC polymer through the photomask, cells attached only to the UV-illuminated region where the MPC polymer was removed, which made the hydrophilic surface relatively hydrophobic. Furthermore, the size of the MC-3T3 E1 cell patterns could be controlled by single cell level. Stability of the cell micropatterns was demonstrated by culturing MC-3T3 E1 cell patterns for 5 weeks on glass slide. The micropatterns were stable during culturing; cell viability also was verified. This method can be a powerful tool for cell patterning research.
本报告描述了一种用于细胞微图案化的新型表面处理技术。利用光化学反应在玻璃表面选择性地控制细胞附着。该策略基于将已知可减少非特异性吸附的2-甲基丙烯酰氧乙基磷酰胆碱(MPC)聚合物与用于选择性细胞图案化的光不稳定连接体(PL)相结合。使用简单的表面改性方法将MPC聚合物直接涂覆在玻璃表面,并通过紫外光照射将其去除。所有表面改性步骤均通过静态水接触角测量、X射线光电子能谱(XPS)、原子力显微镜(AFM)、非特异性蛋白质吸附测量以及细胞附着试验进行评估。通过光掩模对MPC聚合物进行选择性切割后,细胞仅附着于MPC聚合物被去除的紫外光照射区域,该区域使亲水表面相对疏水。此外,MC-3T3 E1细胞图案的尺寸可在单细胞水平上进行控制。通过在载玻片上培养MC-3T3 E1细胞图案5周,证明了细胞微图案的稳定性。在培养过程中微图案稳定;细胞活力也得到了验证。该方法可成为细胞图案化研究的有力工具。