Asakawa Hitoshi, Mochitate Katsumi, Haruyama Tetsuya
Department of Biological Functions and Engineering, Kyushu Institute of Technology, Kitakyushu Science and Research Park, Fukuoka 808-0196, Japan.
Anal Chem. 2008 Mar 1;80(5):1505-11. doi: 10.1021/ac702001u. Epub 2008 Feb 1.
A smart live-cell assay was developed as a cellular biosensing system. This system is based on novel tactics: the direct assembly of human cultured cells onto a cell-adhesive sensing matrix. This novel design provides considerable advantages, among them the possibility of capturing molecular signals immediately after they are secreted from living cells. The design also helps preserve all cellular characteristics intact. In this study, a cell-adhesive NO sensing matrix, acting as both an NO-permeable membrane and a cell-adhesive scaffold, was designed using functional polymers and a short peptide sequence derived from extracellular matrix (ECM) proteins. Using the cell-adhesive NO sensing matrix, we constructed a cellular biosensing system based on in situ monitoring of NO released from a human umbilical vein endothelial cell (HUVEC) layer. HUVECs were employed as an organ-functional model of a blood vessel in view of screening vasodilatory substances for clinical purposes. In our novel system, the electrochemical NO sensor is adjacent to the NO-producing cells, which allows the sensing device to achieve superior sensitivity and precise response to a very low number of NO molecules. Our design enables the fixing of the exact distance between the organ-functional model and the chemical sensor without cumbersome manipulations. Consequently, this cellular biosensing system may be readily applicable to high-throughput analysis in the field of drug screening.
一种智能活细胞检测方法被开发为一种细胞生物传感系统。该系统基于新颖的策略：将人类培养细胞直接组装到细胞粘附传感基质上。这种新颖的设计具有诸多优势，其中包括在分子信号从活细胞分泌后立即捕获它们的可能性。该设计还有助于保持所有细胞特征的完整性。在本研究中，使用功能聚合物和源自细胞外基质（ECM）蛋白的短肽序列设计了一种细胞粘附性NO传感基质，它既作为NO可渗透膜又作为细胞粘附支架。利用该细胞粘附性NO传感基质，我们构建了一个基于原位监测人脐静脉内皮细胞（HUVEC）层释放的NO的细胞生物传感系统。鉴于筛选用于临床目的的血管舒张物质，HUVEC被用作血管的器官功能模型。在我们的新型系统中，电化学NO传感器与产生NO的细胞相邻，这使得传感装置能够对极少量的NO分子实现卓越的灵敏度和精确响应。我们的设计能够在不进行繁琐操作的情况下确定器官功能模型与化学传感器之间的确切距离。因此，这种细胞生物传感系统可能很容易应用于药物筛选领域的高通量分析。
