选择性硅烷去除:培养神经元微图案的技术

J. Corey, B. Wheeler, G. Brewer
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引用次数: 2

摘要

作者开发了一种新的基于光刻胶的技术,用于在玻璃底物上对共价硅烷进行微米分辨率的图图化,以便在低密度培养中对初级海马神经元的生长进行图图化。该技术被命名为选择性硅烷去除,因为底物最初与嗜细胞性氨基硅烷(三甲氧基-硅丙基-二乙烯三胺或DETA)反应,然后通过有图案的光刻胶掩蔽层通过等离子体蚀刻积极去除。该过程是通过将清洗过的表面与憎细胞芳基硅烷(苯基三氯硅烷或PTCS)反应,然后去除光刻胶,冲洗和灭菌来完成的。胚胎大鼠海马神经元以极低密度(20000个细胞/cm/sup 2/)使用无血清B27/Neurobasal(TM)定义的培养基在基质上培养。在这些基质上生长的细胞体中,高达90%的细胞体符合3、5或10 /spl mu/m宽线的网格模式。平均76%的背景方格没有游离的神经突或与图案相连的细胞。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Selective silane removal: a technique for micropatterning of neurons in culture
The authors have developed a new photoresist based technique for micrometer resolution patterning of covalently bound silanes on glassy substrates in order to pattern the growth of primary hippocampal neurons in low density culture. The technique is named Selective Silane Removal because the substrate is initially reacted with a cytophilic aminosilane (trimethoxy-silylpropyl-diethylenetriamine or DETA), which, is then aggressively removed by plasma etching through a masking layer of patterned photoresist. The process is completed by reacting the cleaned surface areas with a cytophobic arylsilane (phenyltrichlorosilane or PTCS) followed by removal of the photoresist, rinsing and sterilization. Embryonic rat hippocampal neurons are cultured on the substrates at very low density (20000 cells/cm/sup 2/) using serum free B27/Neurobasal(TM) defined medium. Up to 90% of cell bodies grown on these substrates complied to a grid pattern of 3, 5 or 10 /spl mu/m wide lines. An average of 76% of the background squares were free of stray neurites or cells connected to the pattern.
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