Hanson Jennifer N, Motala Michael J, Heien Michael L, Gillette Martha, Sweedler Jonathan, Nuzzo Ralph G
Department of Material Science and Engineering, University of Illinois, Urbana-Champaign, Urbana, IL 61801, USA.
Lab Chip. 2009 Jan 7;9(1):122-31. doi: 10.1039/b803595d. Epub 2008 Oct 21.
We explore textural cues as a mechanism for controlling neuronal process outgrowth in primary cultures of mammalian neurons. The work uses a form of decal transfer lithography to generate arrays of PDMS posts of various dimensions and spacings on glass substrates that are rendered growth-compliant by subsequent treatment with a protein activator. Hippocampal neurons plated on these substrates are used to determine how the posts direct process growth by acting as attachment points or guidance cues. Textural features varying over a large range, even as large as 100 microm in diameter, dramatically affect process growth. Indeed, two growth regimes are observed; at the smaller feature sizes considered, process branching strongly aligns (at right angles) along the post mesh, while neuronal outgrowth on the larger feature sizes elicits process wrapping. The latter behavior most strongly manifests in neurons plated initially at approximately 100 cells/mm(2), where the cells were able to form networks, while for isolated neurons, the cells exhibit poorer viability and development. Bag cell neurons from Aplysia californica also display regular growth patterns, but in this case are guided by contact avoidance of the posts, a behavior qualitatively different than that of the hippocampal neurons.
我们探索纹理线索作为控制哺乳动物神经元原代培养中神经突生长的一种机制。这项工作采用一种转印光刻形式,在玻璃基板上生成各种尺寸和间距的聚二甲基硅氧烷柱阵列,通过随后用蛋白质激活剂处理使其具有生长适应性。接种在这些基板上的海马神经元用于确定这些柱子如何作为附着点或引导线索来指导神经突生长。即使直径大到100微米的大范围变化的纹理特征,也会显著影响神经突生长。事实上,观察到两种生长模式;在考虑的较小特征尺寸下,神经突分支沿着柱子网格强烈对齐(成直角),而在较大特征尺寸上的神经元生长则引发神经突缠绕。后一种行为在最初以约100个细胞/mm²接种的神经元中表现最为强烈,在这种情况下细胞能够形成网络,而对于孤立的神经元,细胞表现出较差的活力和发育。来自加州海兔的袋状细胞神经元也表现出规则的生长模式,但在这种情况下是通过避免与柱子接触来引导的,这种行为在性质上与海马神经元不同。