Lee HyungJae, Cho Youngnam
New Experimental Therapeutic Branch, National Cancer Center, 111 Jungbalsan-ro, Ilsandong-gu, Goyang, Gyeonggi-do 410-769, South Korea.
Theranostics. 2015 Jun 10;5(9):1021-9. doi: 10.7150/thno.12389. eCollection 2015.
Here, we report the development of an electric field-assisted methodology for constructing 3D C2C12 cell sheets with the potential for cell surface modification. In this method, a conducting polymer, polypyrrole (Ppy), is electrodeposited via biotin doping, and then chemical conjugation of biotinylated bone morphogenetic protein 2 (BMP2) is achieved using a biotin-streptavidin cross-linker. Subsequently, C2C12 cells are cultured on BMP2-immobilized Ppy surfaces to induce interactions between cell surface receptors and bound BMP2 ligands. Following these procedures, layers of BMP2-immobilized cells can be easily detached from the Ppy surface by applying an electrical potential. This novel method results in high affinity, ligand-bound cell sheets, which exhibit homogeneous coverage with membrane-bound proteins and signal activation that occurs via maximal receptor accessibility. Using this strategy to engineer the cell surface with desirable ligands results in structures that mimic in vivo tissues; thus, the method reported here has potential applications in regenerative medicine and tissue engineering.
在此,我们报告了一种电场辅助方法的开发,该方法用于构建具有细胞表面修饰潜力的三维C2C12细胞片。在该方法中,通过生物素掺杂电沉积导电聚合物聚吡咯(Ppy),然后使用生物素-链霉亲和素交联剂实现生物素化骨形态发生蛋白2(BMP2)的化学偶联。随后,将C2C12细胞培养在固定有BMP2的Ppy表面上,以诱导细胞表面受体与结合的BMP2配体之间的相互作用。按照这些步骤,通过施加电势,可以轻松地将固定有BMP2的细胞层从Ppy表面分离。这种新方法产生了具有高亲和力、配体结合的细胞片,其表现出膜结合蛋白的均匀覆盖以及通过最大受体可及性发生的信号激活。使用这种策略用所需配体对细胞表面进行工程改造会产生模仿体内组织的结构;因此,本文报道的方法在再生医学和组织工程中具有潜在应用。