Takayama Yukiya, Kusamori Kosuke, Hayashi Mika, Tanabe Noriko, Matsuura Satoru, Tsujimura Mari, Katsumi Hidemasa, Sakane Toshiyasu, Nishikawa Makiya, Yamamoto Akira
Department of Biopharmaceutics, Kyoto Pharmaceutical University, Yamashina-ku, Kyoto, 607-8414, Japan.
Laboratory of Biopharmaceutics, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba, 278-8510, Japan.
Sci Rep. 2017 Dec 5;7(1):16953. doi: 10.1038/s41598-017-17166-8.
Mesenchymal stem cells (MSCs) have various functions, making a significant contribution to tissue repair. On the other hand, the viability and function of MSCs are not lasting after an in vivo transplant, and the therapeutic effects of MSCs are limited. Although various chemical modification methods have been applied to MSCs to improve their viability and function, most of conventional drug modification methods are short-term and unstable and cause cytotoxicity. In this study, we developed a method for long-term drug modification to C3H10T1/2 cells, murine mesenchymal stem cells, without any damage, using the avidin-biotin complex method (ABC method). The modification of NanoLuc luciferase (Nluc), a reporter protein, to C3H10T1/2 cells by the ABC method lasted for at least 14 days in vitro without major effects on the cellular characteristics (cell viability, cell proliferation, migration ability, and differentiation ability). Moreover, in vivo, the surface Nluc modification to C3H10T1/2 cells by the ABC method lasted for at least 7 days. Therefore, these results indicate that the ABC method may be useful for long-term surface modification of drugs and for effective MSC-based therapy.
间充质干细胞(MSCs)具有多种功能,对组织修复有重大贡献。另一方面,MSCs在体内移植后的活力和功能并不持久,其治疗效果有限。尽管已将各种化学修饰方法应用于MSCs以提高其活力和功能,但大多数传统的药物修饰方法都是短期且不稳定的,并且会引起细胞毒性。在本研究中,我们使用抗生物素蛋白-生物素复合物方法(ABC法)开发了一种对小鼠间充质干细胞C3H10T1/2细胞进行长期药物修饰的方法,且不会造成任何损伤。通过ABC法将报告蛋白纳米荧光素酶(Nluc)修饰到C3H10T1/2细胞上,在体外至少持续14天,对细胞特性(细胞活力、细胞增殖、迁移能力和分化能力)没有重大影响。此外,在体内,通过ABC法对C3H10T1/2细胞进行的表面Nluc修饰至少持续7天。因此,这些结果表明ABC法可能有助于药物的长期表面修饰以及基于MSCs的有效治疗。