自组装IKVAV多肽纳米纤维支架凝胶及其对嗅鞘细胞的作用.pdf

生 物 工 程 学 报 Chin J Biotech 2009, February 25; 25(2): 292-298 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ © 2009 Institute of Microbiology, CAS CSM, All rights reserved 组织工程与细胞培养 自组装IKVAV 多肽纳米纤维支架凝胶及其对嗅鞘细胞 的作用 徐会法, 邵增务, 吴永超, 邓超, 俞旭东, 丁凡, 张保龙, 徐蔚蔚 华中科技大学同济医学院附属协和医院骨科, 武汉 430022 摘 要: 旨在观察自组装IKVAV 多肽纳米纤维支架凝胶对鼠嗅鞘细胞(OECs)的作用。通过调整IKVAV 溶液pH 值并加 入培养液触发多肽自组装为支架凝胶, 用原子力显微镜检测IKVAV 分子可以自组装成编织状纳米纤维(直径为3~5 nm)。 采用原代分离培养方法获得OECs 单细胞悬液后, 使用差速贴壁法两次纯化OECs 且在第 12 天通过免疫染色计数OECs 纯度为 85%。将IKVAV 多肽纳米纤维支架凝胶与OECs 复合培养, 倒置显微镜下观察 OECs 生长良好, Calcein-AM/PI 活、死细胞染色表明活细胞数达95% 。CCK-8 法间接细胞计数证实IKVAV 多肽可促进OECs 的黏附, 对 OECs 增殖没 有影响。由此可见 IKVAV 多肽可以自组装成纳米纤维支架凝胶且对OECs 有良好的生物相容性及黏附作用, 可作为神 经组织工程支架材料。 关键词: 纳米纤维, 组织工程, 自组装, 嗅鞘细胞 Effects of self-assembled IKVAV peptide nanofibers on olfactory ensheathing cells Huifa Xu, Zengwu Shao, Yongchao Wu, Chao Deng, Xudong Yu, Fan Ding, Baolong Zhang, and Weiwei Xu Department of Orthopaedics , Union Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430022, China Abstract: We observed the effects of IKVAV self-assembling peptide nanofiber scaffold (SAPNS) on olfactory ensheathing cells (OECs). The IKVAV molecules were triggered to self- assemble to interconnected nanofibers hydrogel by adjusting pH of solution and adding of DMEM/F12 culture medium. Atomic Force Microscopy (AFM) showed that self-assembly hydrogel was consisted of the interconnected nanofibers, which varied from three nm to five nm in diameter and hundreds nanometer in length. The primary OECs were isolated from rat olfactory