We developed novel scaffolds for the adherent culture of neural stem/progenitor cells on the woven mesh. Nylon mesh (NM) is an inert material for cell adhesion. We prepared polyacrylic acid-grafted nylon mesh (PAA-NM) by graft polymerization method using gamma-irradiation. Matrigel was covalently immobilized to the carboxyl groups in PAA-NM by chemical conjugation using 1-ethyl-3-(3-dimethylamino propyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to prepare the Matrigel-immobilized PAA-grafted nylon mesh (M-PAA-NM). Cell adhesion property of mouse neural stem/progenitor cells (NSPCs) between the NM, PAA-NM, and M-PAA-NM was different from each other. The neurosphere-like clusters of NSPCs were weakly bound to NM and PAA-NM without spreading. The NSPCs were firmly adhered to, spread, and covered the surface of M-PAA-NM. We evaluated the state of differentiation by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immnocytochemistry. A neuronal marker β III tubulin, a glial marker glial fibrillary acidic protein (GFAP) and a mature glial marker S100β were expressed at a low level in the cultured cells while immature NSPCs marker Nestin and Sox2 were slightly lower without significant statistical difference. We concluded that the M-PAA-NM is a good substrate for adherent culture of NSPCs without triggering their cell differentiation, and also provides the maintenance of their growth with fewer passages in comparison with the conventional suspension culture of NSPCs in neurospheres.

我们开发了新型支架，用于在编织网上贴壁培养神经干/祖细胞。尼龙网（NM）是一种用于细胞粘附的惰性材料。我们通过使用γ射线的接枝聚合方法制备了聚丙烯酸接枝尼龙网（PAA-NM）。通过使用1-乙基-3-（3-二甲基氨基丙基）碳二亚胺（EDC）和N进行化学共轭，将Matrigel共价固定在PAA-NM中的羧基上-羟基琥珀酰亚胺（NHS）制备基质胶固定的PAA接枝尼龙网（M-PAA-NM）。NM，PAA-NM和M-PAA-NM之间的小鼠神经干/祖细胞（NSPC）的细胞粘附特性彼此不同。NSPCs的神经球样簇弱结合到NM和PAA-NM而不扩散。NSPC牢固地粘附，扩散并覆盖了M-PAA-NM的表面。我们通过定量逆转录聚合酶链反应（RT-PCR）和免疫细胞化学评估了分化状态。神经元标志物βIII微管蛋白，神经胶质标志物胶质纤维酸性蛋白（GFAP）和成熟神经胶质标志物S100β在培养细胞中低水平表达，而未成熟的NSPCs标志物Nestin和Sox2则略低，而无统计学差异。