Development of an artificial neuronal network with post-mitotic rat fetal hippocampal cells by polyethylenimine
Development of an artificial neuronal network with post-mitotic rat fetal hippocampal cells by polyethylenimine
复制标题
用聚乙烯亚胺开发有丝分裂后大鼠胎儿海马细胞的人工神经元网络
DOI：
10.1016/j.bios.2007.11.007
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发表时间：
2008-03-14
影响因子：
12.6
通讯作者：
Xu, Qunyuan
中科院分区：
文献类型：
--
作者：
Liu, Bingfang;Ma, Jun;Xu, Qunyuan
在设计简化的体外神经元网络时，选择合适的表面材料促进细胞的黏附和生长是一个重要的考虑因素。过去，细胞外基质蛋白如层粘连蛋白(LN)或正电荷物质如多聚L赖氨酸(PLL)已被使用。在这项研究中，我们检测了另一种带正电的聚合物，聚乙烯亚胺(PEI)，在体外促进神经元黏附、生长和形成功能性神经元网络的能力。以PEI、PLL和LN为原料，采用微接触印刷的方法在玻片上制作网格状图案。将大鼠胚胎海马区有丝分裂后神经元在不同聚合物上培养，在无血清培养条件下，用荧光显微镜和原子力显微镜(AFM)观察神经元的存活率和形态。我们发现，培养在PEI和PLL涂层表面的神经元沿着网格状图案附着和延伸，而培养在LN涂层盖片上的神经元聚集成细胞团。此外，我们还发现，PEI和PLL涂层网格上的神经元在无血清条件下存活了2周以上，而LN涂层网格上培养的神经元大部分在1周后死亡。利用原子力显微镜，我们观察到在PEI包被的覆盖物的神经元胞体附近有一些神经突触样结构。这些发现表明PEI是在体外建立功能性神经元网络的合适表面。(C)2007年，爱思唯尔出版。
The selection of appropriate surface materials that promote cellular adhesion and growth is an important consideration when designing a simplified neuronal network in vitro. In the past, extracellular matrix proteins such as laminin (LN) or positively charged substances such as POIY-L-lysine (PLL) have been used. In this study, we examined the ability of another positively charged polymer, polyethyleneimine (PEI), to promote neuronal adhesion, growth and the formation of a functional neuronal network in vitro. PEI, PLL and LN were used to produce grid-shape patterns on glass coverslips by micro-contact printing. Post-mitotic neurons from the rat fetal hippocampus were cultured on the different polymers and the viability and morphology of these neurons under serum-free culture conditions were observed using fluorescent microscopy and atomic force microscopy (AFM). We show that neurons cultured on the PEI- and PLL-coated surfaces adhered to and extended neurites along the grid-shape patterns, whereas neurons cultured on the LN-coated coverslips clustered into clumps of cells. In addition, we found that the neurons on the PEI and PLL-coated grids survived for more than 2 weeks in serum-free conditions, whereas most neurons cultured on the LN-coated grids died after 1 week. Using AFM, we observed some neurosynapse-like structures near the neuronal soma on PEI-coated coverstips. These findings indicate that PEI is a suitable surface for establishing a functional neuronal network in vitro. (c) 2007 Published by Elsevier B.V.