神经干细胞与功能化自组装多肽水凝胶的细胞相容性

目的 制备功能化自组装多肽水凝胶并检测其与神经干细胞(NSCs)的生物相容性.方法 合成自组装多肽RADA16 与多肽RADA16-IKVAV,将两者混合制备功能化自组装多肽水凝胶,用原子力显微镜(AFM)观察其形态学特征.采用无血清培养法从新生大鼠皮层分离培养NSCs.将NSCs分别接种到RADA16自组装多肽水凝胶(对照组)与功能化自组装多肽水凝胶(实验组)表面.激光扫描共聚焦显微镜(LSCM)观察细胞迁移.用CCK-8法测定吸光度(A)检测细胞增殖能力.应用Nestin、MAP2、GFAP和CC-1免疫荧光染色,检测神经干细胞分化.结果 AFM显示功能化自组装多肽水凝胶由纳米纤维组成,其纤维直径为20～30 nm,长度可达数百纳米.在细胞接种6 d后,对照组和实验组细胞在水凝胶中的迁移距离分别为(27.5±3.7)μm和(53.2±5.4)μm,两组差异有统计学意义(P＜0.05).复合培养7 d后,实验组中细胞A值明显高于对照组(P＜0.05).免疫荧光结果显示13 d后对照组中MAP2阳性细胞百分率为(22.44±3.52)%,实验组为(40.35±4.84)%,两组差异有统计学意义(P＜0.05).结论 功能化自组装多肽水凝胶与NSCs相容性良好.

Cytocompatibility of functionalized self-assembling peptide hydrogel with neural stem cells

Objective To develop a functionalized self-assembling peptide hydrogel.and investigate its biocompatibility with neural stem cells(NSCs).Methods The functionalized self-assembling peptide was made by mixing the self-assembling peptide RADA16 and peptide RADA16-IKVAV.The norphologieal features of the functionalized self-assembling peptide were studied by atom force microscope (AFM).NSCs were isolated from the cerebral cortex of neonatal rats and cultured in serum-free medium.NSCs were seeded on the surface of both RADA16 self-assembling peptide hydrogel(control group)and the funetionalized self-assembling peptide hydrogel(experimental group).The proliferation behavior of the cells was evaluated by CCK-8 assay as expressed by absorbance value.Cell migration was detected by laser scan confocal microscope(LsCM).Immunofluorescenee staining with Nestin,MAP2,GFAP,and CC-1 was used to assess the differentiation of NSCs.Results AFM showed that the functionalized self-assembling peptide hydrogel was made up of nanofibers with diameter of 20-30 nm and length of hundreds nanometers.In 6 days after cell seeding,the depth of cell migration in experimental group(53.2±5.4)μm]was significantly higher than that in control group(27.5 4-3.7)μm,P＜0.05].CCK-8 assay showed that experimental group displayed a significant increase of absorbance value compared with that of the control group after 1-week culture(P＜0.05).Immunofluorenscene analysis indicated that the differentiation ratio of neurons from NSCs in control group and experimental group were(22.44±3.52)% and(40.35±4.84)%respectively after 13-days cultare.The difference between the two groups was significant(P＜0.05).Conclusion The functionalized self-assembling peptide hydrogel has good cytocompatibility with NSCs.