An efficient surface modification using 2-methacryloyloxyethyl phosphorylcholine to control cell attachment via photochemical reaction in a microchannel
An efficient surface modification using 2-methacryloyloxyethyl phosphorylcholine to control cell attachment via photochemical reaction in a microchannel
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DOI：
10.1039/c002239j
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Kitamori, Takehiko
中科院分区：
文献类型：
--
作者：
Jang, Kihoon;Sato, Kae;Kitamori, Takehiko
该报告描述了一种在封闭玻璃微通道中进行细胞微图案化的直接方法。为了控制微通道内的细胞粘附性，外部刺激（例如紫外线（UV））的应用是必不可少的。该技术侧重于使用改性 2-甲基丙烯酰氧基乙基磷酰胆碱 (MPC) 聚合物，已知该聚合物是一种非生物污染化合物，是一种光可裂解连接体 (PL)，通过连接到氨基封端的硅烷化表面来定位细胞。使用紫外光照射，MPC 聚合物通过控制微通道内细胞附着的光化学反应被选择性消除。为了在微通道中形成合适的细胞微图案，研究了细胞悬浮液的最佳紫外线照射时间和浓度。通过光掩模选择性去除 MPC 聚合物后，MC-3T3 E1 细胞和血管内皮细胞 (EC) 仅定位于紫外线暴露区域。此外，通过在流动条件下在微通道中培养2周也证实了图案化EC的稳定性。此外，我们通过多次去除 MPC 聚合物，在同一微通道内使用了两种不同类型的细胞。 EC 和 Piccell 分别位于微通道的上游和下游。当内皮细胞受到三磷酸腺苷（ATP）刺激时，内皮细胞会分泌一氧化氮，并且可以通过Piccells中的荧光共振能量转移（FRET）来检测，这是一种基于细胞的一氧化氮指示剂。该技术可以成为分析细胞相互作用研究的强大工具。
This report describes a direct approach for cell micropatterning in a closed glass microchannel. To control the cell adhesiveness inside the microchannel, the application of an external stimulus such as ultraviolet (UV) was indispensible. This technique focused on the use of a modified 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which is known to be a non-biofouling compound that is a photocleavable linker (PL), to localize cells via connection to an amino-terminated silanized surface. Using UV light illumination, the MPC polymer was selectively eliminated by photochemical reaction that controlled the cell attachment inside the microchannel. For suitable cell micropatterning in a microchannel, the optimal UV illumination time and concentration for cell suspension were investigated. After selective removal of the MPC polymer through the photomask, MC-3T3 E1 cells and vascular endothelial cells (ECs) were localized only to the UV-exposed area. In addition, the stability of patterned ECs was also confirmed by culturing for 2 weeks in a microchannel under flow conditions. Furthermore, we employed two different types of cells inside the same microchannel through multiple removal of the MPC polymer. ECs and Piccells were localized in both the upper and down streams of the microchannel, respectively. When the ECs were stimulated by adenosine triphosphate (ATP), NO was secreted from the ECs and could be detected by fluorescence resonance energy transfer (FRET) in Piccells, which is a cell-based NO indicator. This technique can be a powerful tool for analyzing cell interaction research.