Single-cell attachment and culture method using a photochemical reaction in a closed microfluidic system
Single-cell attachment and culture method using a photochemical reaction in a closed microfluidic system
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DOI：
10.1063/1.3494287
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发表时间：
2010-09-01
期刊：
影响因子：
3.2
通讯作者：
Kitamori, Takehiko
中科院分区：
文献类型：
--
作者：
Jang, Kihoon;Xu, Yan;Kitamori, Takehiko
最近，人们对单细胞分析的兴趣有所增加，因为它有可能提高我们对细胞过程的理解。单细胞操作和贴壁对于实现这一任务是必不可少的。在本文中，我们采用了一种简单直接的方法在封闭的微通道中进行单细胞附着和培养。通过应用非生物污染聚合物、2-甲基丙烯酰氧基乙基磷酰胆碱（MPC）聚合物和硝基苄基光可裂解连接体对微通道表面进行改性。使用紫外 (UV) 光照射，通过光化学反应选择性去除 MPC 聚合物，从而调节微通道内的细胞粘附。为了在微通道中获得所需的单个内皮细胞图案，通过使用直径为10、20、30或50μm的圆形光掩模来控制细胞粘附区域。仅当使用20和30μm光掩模时，孵育12小时后形成单细胞贴壁图案，并且贴壁和非贴壁细胞在整个紫外照射区域中的比例分别为21.3％+/-0.3％和7.9％+/-0.3％。 20μm光掩模情况下的单细胞粘附频率是30μm光掩模情况下的2.7倍。我们发现 20 μm 光掩模最适合在微通道中形成单细胞粘附图案。该技术可以成为分析细胞表面和细胞-细胞外基质接触等环境因素的强大工具。 (C) 2010 年美国物理研究所。 [doi:10.1063/1.3494287]
Recently, interest in single cell analysis has increased because of its potential for improving our understanding of cellular processes. Single cell operation and attachment is indispensable to realize this task. In this paper, we employed a simple and direct method for single-cell attachment and culture in a closed microchannel. The microchannel surface was modified by applying a nonbiofouling polymer, 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, and a nitrobenzyl photocleavable linker. Using ultraviolet (UV) light irradiation, the MPC polymer was selectively removed by a photochemical reaction that adjusted the cell adherence inside the microchannel. To obtain the desired single endothelial cell patterning in the microchannel, cell-adhesive regions were controlled by use of round photomasks with diameters of 10, 20, 30, or 50 mu m. Single-cell adherence patterns were formed after 12 h of incubation, only when 20 and 30 mu m photomasks were used, and the proportions of adherent and nonadherent cells among the entire UV-illuminated areas were 21.3%+/- 0.3% and 7.9%+/- 0.3%, respectively. The frequency of single-cell adherence in the case of the 20 mu m photomask was 2.7 times greater than that in the case of the 30 mu m photomask. We found that the 20 mu m photomask was optimal for the formation of single-cell adherence patterns in the microchannel. This technique can be a powerful tool for analyzing environmental factors like cell-surface and cell-extracellular matrix contact. (C) 2010 American Institute of Physics. [doi:10.1063/1.3494287]