Novel high-resolution micropatterning for neuron culture using polylysine adsorption on a cell repellant, plasma-polymerized background.
Novel high-resolution micropatterning for neuron culture using polylysine adsorption on a cell repellant, plasma-polymerized background.
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DOI：
10.1021/la8021479
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发表时间：
2008-11-18
期刊：
影响因子：
--
通讯作者：
Sretavan DW
中科院分区：
文献类型：
--
作者：
Chang WC;Sretavan DW
在培养中组织单个神经元及其过程的能力为基础神经科学研究应用和生物医学微设备的发展提供了重要的好处。虽然已经有许多方法被用于产生神经元和细胞的微图案，但目前还没有方法可以同时提供高分辨率的图案，同时使细胞对所需图案的高度依从性和良好的可制造性。为了开发这样的工艺，这项工作使用了等离子体聚合，无污染的聚环氧乙烷（PEO）样膜来提供细胞排斥基底，细胞粘附微图案可以选择性地放置在其上。虽然使用等离子体聚合的有机膜已被用于细胞微图图化，但该工艺利用了这种peo样材料表面经常被忽视的倾向，即从水溶液中吸附聚赖氨酸，同时保持对其他物质（如牛血清白蛋白（BSA）和免疫球蛋白G (IgG)）的不污染。当聚赖氨酸的吸附被短暂的等离子体氧化增强时，材料的表面化学性质会发生轻微的变化，简单的光刻提升可以用来在细胞排斥的背景上形成稳定的细胞粘附区域的微图案。我们表明，光刻本身在peo类材料上的应用不会改变其化学性质，也不会导致其表面微图案聚赖氨酸的侵蚀。来自胚胎小鼠的海马神经元在这些微图案基质上繁殖，并表现出与在聚赖氨酸涂层玻璃上培养的神经元相当的活力。此外，细胞体和离体神经突对微图案的顺应性几乎是完美的。除了提供细胞粘附区域外，微图型聚赖氨酸涂层还作为模板介导其他生物活性物质（如IgG和层粘连蛋白）的固定化。利用聚赖氨酸上的层粘连蛋白，我们还能够培养和微模式视网膜神经节细胞（RGC）。
The ability to organize individual neurons and their processes in culture provides important benefits to both basic neuroscience research applications and the development of biomedical microdevices. While numerous methods have been used to produce such micropatterning of neurons and cells in general, there has yet been no method to simultaneously provide high-resolution patterns with high compliance of cells to desired patterns and good manufacturability. To develop such a process, this work used a plasma polymerized, nonfouling poly ethylene oxide (PEO)-like film to provide a cell repellant substrate on which cell adhesive micropatterns can be selectively laid down. While the use of plasma polymerized, organic films have been used for cell micropatterning, this process exploits the often-overlooked tendency for the surface of this PEO-like material to adsorb polylysine from aqueous solution while remaining nonfouling with respect to other species, such as bovine serum albumin (BSA) and immunoglobulin G (IgG). When the adsorption of polylysine was enhanced by brief plasma oxidation, which slightly alters the surface chemistry of the material, simple photolithographic liftoff could be used to micropattern stable, cell adhesive areas on an otherwise cell repellant background. We showed that the application of photolithography itself on the PEO-like material did not alter its chemical properties, nor did it result in the erosion of the micropatterned polylysine on its surface. Hippocampal neurons from embryonic mice flourished on these micropatterned substrates and exhibited viability comparable to neurons cultured on polylysine coated glass. Furthermore, the compliance of cell bodies and outgrowing neurites to the micropatterns was nearly perfect. In addition to providing cell adhesive regions, the micropatterned polylysine coating also served as a template mediating the immobilization of other bioactive species such as IgG and laminin. Using this “piggybacking” of laminin on polylysine, we were also able to culture and micropattern retinal ganglion cells (RGC).
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