Yamamoto Hideaki, Demura Takanori, Sekine Kohei, Kono Sho, Niwano Michio, Hirano-Iwata Ayumi, Tanii Takashi
Frontier Research Institute for Interdisciplinary Sciences, Tohoku University; CREST, Japan Science and Technology Agency;
School of Fundamental Science and Engineering, Waseda University.
J Vis Exp. 2015 Oct 26(105):e53045. doi: 10.3791/53045.
Organic contaminants adsorbed on the surface of titanium dioxide (TiO2) can be decomposed by photocatalysis under ultraviolet (UV) light. Here we describe a novel protocol employing the TiO2 photocatalysis to locally alter cell affinity of the substrate surface. For this experiment, a thin TiO2 film was sputter-coated on a glass coverslip, and the TiO2 surface was subsequently modified with an organosilane monolayer derived from octadecyltrichlorosilane (OTS), which inhibits cell adhesion. The sample was immersed in a cell culture medium, and focused UV light was irradiated to an octagonal region. When a neuronal cell line PC12 cells were plated on the sample, cells adhered only on the UV-irradiated area. We further show that this surface modification can also be performed in situ, i.e., even when cells are growing on the substrate. Proper modification of the surface required an extracellular matrix protein collagen to be present in the medium at the time of UV irradiation. The technique presented here can potentially be employed in patterning multiple cell types for constructing coculture systems or to arbitrarily manipulate cells under culture.
吸附在二氧化钛(TiO₂)表面的有机污染物可在紫外(UV)光下通过光催化作用分解。在此,我们描述了一种采用TiO₂光催化作用来局部改变底物表面细胞亲和力的新方法。对于本实验,在玻璃盖玻片上溅射涂覆一层TiO₂薄膜,随后用源自十八烷基三氯硅烷(OTS)的有机硅烷单分子层对TiO₂表面进行修饰,该单分子层会抑制细胞黏附。将样品浸入细胞培养基中,并将聚焦的紫外光照射到一个八边形区域。当将神经母细胞瘤细胞系PC12细胞接种到样品上时,细胞仅黏附在紫外照射区域。我们进一步表明,这种表面修饰也可以原位进行,即便是细胞在底物上生长时。表面的适当修饰要求在紫外照射时培养基中存在细胞外基质蛋白胶原蛋白。这里介绍的技术有可能用于对多种细胞类型进行图案化处理,以构建共培养系统,或者在培养过程中任意操控细胞。