Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer
Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer
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DOI：
10.1016/s1385-299x(00)00024-6
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发表时间：
2000-07-01
期刊：
影响因子：
--
通讯作者：
Linial, M
中科院分区：
文献类型：
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作者：
Bledi, Y;Domb, AJ;Linial, M
在培养中维持细胞对于研究细胞生物学和生理学的许多方面是必不可少的。细胞培养依赖于细胞在生长表面的适当锚定。对于大多数细胞系，特别是有丝分裂后的神经元，包被的组织培养板是接种的先决条件。最常用的涂层试剂是带正电荷的聚合物，如聚-L-赖氨酸或生物纯化的粘附分子，如胶原蛋白。在这份报告中，我们提出了一个简单的程序合成和涂层细胞培养表面。该试剂是一种生物惰性的疏水化聚乙烯亚胺（PEI），它为培养的细胞系（包括神经元谱系的细胞系）提供了足够的粘附性能。疏水化的PEI是通过与PEI的2摩尔%的氨基结合的十八烷基改性的支链PEI。与水溶性的天然PEI不同，改性PEI可溶于乙醇，因此在生物介质中耐溶解。对组织培养板和玻璃表面的包被方案进行了优化，在许多方面，该聚合物优于其他常规使用的包被试剂。接种在聚合物处理的表面上的神经元细胞系对包括反复更换培养基和大量洗涤在内的操作具有抗性。与其他常用的涂层试剂相比，开发的PEI基聚合物涂层表面的优点进行了讨论。(C)2000 Elsevier Science B.V.保留所有权利。
Maintaining cells in culture is essential for studying many aspects of cell biology and physiology. Cell culturing is dependent on proper anchorage of cells to the growth surfaces. For most cell lines, and especially for post-mitotic neurons, coated tissue culture plates are prerequisite for seeding. The most commonly used coating reagents are positively charged polymers such as poly-L-lysine or biologically purified adhesive molecules such as collagen. In this report, we present a simple procedure for synthesizing and for coating cell culture surfaces. The reagent is a biologically inert hydrophobized polyethyleneimine (PEI), which provides adequate adhesive properties for cultured cell lines including those of neuronal lineage. The hydrophobized PEI is branched PEI modified by octadecanyl groups bound to 2 mol% of the amino groups of the PEI. Unlike the native PEI that is water soluble, the modified PEI is soluble in ethanol, and thus resistant to solubilization in biological media. The protocol of coating was optimized for tissue culture plates as well as glass surfaces and in many respects this polymer outperformed other routinely used coating reagents. Neuronal cell lines, plated on the polymer-treated surfaces are resistant to manipulations including repeated media changes and extensive washing. The advantage of coating surfaces with the developed PEI-based polymer compared to other commonly used coating reagents is discussed. (C) 2000 Elsevier Science B.V. All rights reserved.