Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation
Combinatorial protein display for the cell-based screening of biomaterials that direct neural stem cell differentiation
复制标题
DOI：
10.1016/j.biomaterials.2006.10.004
复制
发表时间：
2007-02-01
期刊：
影响因子：
14
通讯作者：
Iwata, Hiroo
中科院分区：
文献类型：
--
作者：
Nakajima, Masafumi;Ishimuro, Toshinari;Iwata, Hiroo
神经干细胞（NSC）已成为创伤性损伤和中枢神经系统退行性疾病后细胞替代治疗的潜在来源。然而，NSC的临床应用还需要技术进步，特别是用于控制NSC的分化。本研究旨在开发用于扩增未分化NSC或诱导具有特定表型的细胞的生物材料。我们的方法是构建由细胞外基质成分和生长因子组成的复合生物材料。为了优化基质-生长因子组合，我们通过使用基于细胞的阵列的测定进行了复合生物材料的平行和快速筛选。光辅助图案化的烷乙醇胺自组装单分子层，以实现与生长因子，包括表皮生长因子（EGF），睫状神经营养因子（CNTF），神经生长因子（NGF），和神经营养因子-3（NT-3）的天然和合成的基质掺入位点可寻址的组合固定。从大鼠胚胎纹状体获得的NSC直接培养在阵列上筛选细胞粘附、增殖以及促进神经元和胶质细胞的特化。结果表明，大量细胞粘附于层粘连蛋白-1、纤连蛋白、ProNectin（TM）和聚（乙烯亚胺）。结果发现，细胞增殖最广泛的一个点与固定的EGF之间的斑点与不同的基质生长因子的组合。结果还表明，固定化NGF或NT-3促进神经元分化，CNTF促进星形胶质细胞分化。重要的是，观察到的生长因子的影响经常改变取决于类型的共固定化矩阵，这表明粘附和生长因子信号的协同效应。(c)2006爱思唯尔有限公司保留所有权利。
Neural stem cell (NSC) has emerged as a potential source for cell replacement therapy following traumatic injuries and degenerative diseases of the central nervous system. However, clinical applications of NSC further require technological advances especially for controlling differentiation of NSC. This study aimed at developing biomaterials that serve to expand undifferentiated NSC or to induce cells with specific phenotypes. Our approach is to construct composite biomaterials that consist of extracellular matrix components and growth factors. In order to optimize matrix-growth factor combinations, we conducted the parallel and rapid screening of composite biomaterials through assays using cell-based arrays. The photo-assisted patterning of an alkanethiol self-assembled monolayer was employed to achieve site-addressable combinatorial immobilization of natural and synthetic matrices incorporated with growth factors including epidermal growth factor (EGF), ciliary neurotrophic factor (CNTF), nerve growth factor (NGF), and neurotrophin-3 (NT-3). NSC obtained from the rat embryonic striatum was cultured directly on the array to screen for cell adhesion, proliferation, and promotion of neuronal and glial specification. The results showed that the significant number of cells adhered to laminin-1, fibronectin, ProNectin (TM), and poly(ethyleneimine). It was found that cells proliferated most extensively on a spot with immobilized EGF among the spots with different matrix-growth factor combinations. The results also showed that neuronal differentiation was promoted on the spots with immobilized NGF or NT-3, and astroglial differentiation with CNTF. Importantly, observed effects of growth factors were frequently altered depending on the type of co-immobilized matrices, suggesting synergic effects of adhesion and growth factor signals. (c) 2006 Elsevier Ltd. All rights reserved.