Long-term stability of grafted polyethylene glycol surfaces for use with microstamped substrates in neuronal cell culture
Long-term stability of grafted polyethylene glycol surfaces for use with microstamped substrates in neuronal cell culture
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DOI：
10.1016/s0142-9612(00)00343-4
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发表时间：
2001-05-01
期刊：
影响因子：
14
通讯作者：
Leckband, DE
中科院分区：
文献类型：
--
作者：
Branch, DW;Wheeler, BC;Leckband, DE
对于神经元微模式的长期稳定至关重要的是在暴露于细胞培养条件下底层的功能保留。我们报道了共价结合的PEG膜在长期细胞培养中持续延缓蛋白质粘附和细胞生长的能力。PDMS微印章用于制造允许细胞附着和生长的聚d-赖氨酸（PDL）底物，聚乙二醇（PEG）底物用于减少蛋白质和细胞粘附。利用零椭偏仪和原子力显微镜（AFM）测量薄膜厚度。利用傅里叶变换红外光谱（FT-IR）对有机硅烷薄膜结构进行了研究。在0.1 M磷酸钠缓冲液pH 7.4中浸泡长达一个月，测试细胞培养条件下的长期膜稳定性。零椭偏测量和水接触测量表明，有机硅烷膜在一个月内保持稳定，而聚乙二醇膜厚度在25天后迅速下降。在均匀的PEG底物上以200个细胞/毫米(2)的速度涂覆的海马细胞在整个培养过程中，PEG膜上的生物膜厚度稳步增加，可能来自神经元来源的蛋白质。我们发现，在细胞培养条件下，交联过程中的所有层都是稳定的，除了PEG在25天后降解。2001爱思唯尔科学有限公司版权所有。
Crucial to long-term stability of neuronal micropatterns is functional retention of the underlying substratum while exposed to cell culture conditions. We report on the ability of covalently bound PEG films in long-term cell culture to continually retard protein adhesion and cell growth. PDMS microstamps were used to create poly-d-lysine (PDL) substrates permissive to cell attachment and growth, and polyethylene glycol(PEG) substrates were used to minimize protein and cell adhesion. Film thickness was measured using null ellipsometry and atomic force microscopy (AFM). Organosilane film structure was examined using Fourier transform infrared (FT-IR) spectroscopy. Long-term film stability in cell culture conditions was tested by immersion in 0.1 M sodium phosphate buffer pH 7.4 for up to one month. Null ellipsometry and water contact measurements indicated that organosilane films were stable up to one month, whereas the PEG film thickness declined rapidly after day 25. Hippocampal cells plated at 200 cells/mm(2) on uniform PEG substrates gave a steady increase in biofilm thickness on PEG films throughout the culture, possibly from proteins of neuronal origin. We found that all the layers in the cross-linking procedure were stable in cell culture conditions, with the exception of PEG, which degraded after day 25. (C) 2001 Elsevier Science Ltd. All rights reserved.