Graduate School of Pure and Applied Sciences, University of Tsukuba, Ibaraki, Japan.
Tissue Eng Part A. 2013 Jan;19(1-2):290-8. doi: 10.1089/ten.TEA.2011.0739. Epub 2012 Sep 5.
In this study, we describe the development of oligopeptide-modified cell culture surfaces from which adherent cells can be rapidly detached by application of an electrical stimulus. An oligopeptide, CGGGKEKEKEK, was designed with a terminal cysteine residue to mediate binding to a gold surface via a gold-thiolate bond. The peptide forms a self-assembled monolayer through the electrostatic force between the sequence of alternating charged glutamic acid (E) and lysine (K) residues. The dense and electrically neutral oligopeptide zwitterionic layer of the modified surface was resistant to nonspecific adsorption of proteins and adhesion of cells, while the surface was altered to cell adhesive by the addition of a second oligopeptide (CGGGKEKEKEKGRGDSP) containing the RGD cell adhesion motif. Application of a negative electrical potential to this gold surface cleaved the gold-thiolate bond, leading to desorption of the oligopeptide layer, and rapid (within 2 min) detachment of virtually all cells. This approach was applicable not only to detachment of cell sheets but also for transfer of cell micropatterns to a hydrogel. This electrochemical approach of cell detachment may be a useful tool for tissue-engineering applications.
在这项研究中,我们描述了一种由短肽修饰的细胞培养表面的开发,通过施加电刺激可以快速将贴壁细胞从该表面上脱离。我们设计了一个带有末端半胱氨酸残基的短肽 CGGGKEKEKEK,以通过金-硫醇键介导与金表面的结合。该短肽通过序列中交替的带电荷的谷氨酸(E)和赖氨酸(K)残基之间的静电力形成自组装单层。修饰表面的密集且电中性的短肽两性离子层能抵抗蛋白质的非特异性吸附和细胞的黏附,而通过添加包含 RGD 细胞黏附基序的第二个短肽(CGGGKEKEKEKGRGDSP),表面又能变成黏附细胞的状态。施加负的电势会导致金-硫醇键断裂,从而使短肽层解吸,并迅速(在 2 分钟内)几乎将所有细胞脱离。这种电化学细胞脱离方法不仅适用于细胞片的脱离,也适用于细胞微图案向水凝胶的转移。这种电化学细胞脱离方法可能是组织工程应用的有用工具。