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通过微通道中的光化学反应,使用 2-甲基丙烯酰氧乙基磷酸胆碱对表面进行高效改性,从而控制细胞附着。

An efficient surface modification using 2-methacryloyloxyethyl phosphorylcholine to control cell attachment via photochemical reaction in a microchannel.

机构信息

Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.

出版信息

Lab Chip. 2010 Aug 7;10(15):1937-45. doi: 10.1039/c002239j. Epub 2010 May 25.

Abstract

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.

摘要

本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。为了控制微通道内细胞的粘附力,需要应用外部刺激,如紫外线 (UV)。该技术专注于使用改性的 2-(甲基丙烯酰氧基)乙基磷酸胆碱 (MPC) 聚合物,该聚合物是一种已知的非生物污染化合物,也是一种光可裂解的连接剂 (PL),通过连接到氨基封端的硅烷化表面来定位细胞。通过光化学反应,MPC 聚合物可以被选择性地消除,从而控制微通道内细胞的附着。为了在微通道中进行合适的细胞微图案化,研究了细胞悬浮液的最佳 UV 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) 仅被定位到 UV 暴露区域。此外,通过在微通道中在流动条件下培养 2 周,还证实了图案化 EC 的稳定性。此外,我们通过多次去除 MPC 聚合物,在同一个微通道中使用两种不同类型的细胞。EC 和 Piccells 分别定位在微通道的上游和下游。当 EC 受到三磷酸腺苷 (ATP) 的刺激时,EC 会分泌一氧化氮 (NO),Piccells 可以通过荧光共振能量转移 (FRET) 检测到,Piccells 是一种基于细胞的 NO 指示剂。该技术可以成为分析细胞相互作用研究的有力工具。

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