Directed cell growth on protein-functionalized hydrogel surfaces
Directed cell growth on protein-functionalized hydrogel surfaces
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DOI：
10.1016/j.jneumeth.2007.01.024
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发表时间：
2007-05-15
影响因子：
3
通讯作者：
Shain, William
中科院分区：
文献类型：
--
作者：
Hynd, Matthew R.;Frampton, John P.;Shain, William
生物化学表面修饰已用于指导细胞在生物相容性凝胶表面上的附着和生长。在丙烯酰-链霉亲和素单体的存在下光聚合丙烯酰胺基水凝胶，以产生能够结合生物素标记的蛋白质的平面的功能化表面。使用蛋白质的软蛋白质光刻（微接触印刷）将生物素化的细胞外基质蛋白质、纤连蛋白和层粘连蛋白以及层粘连蛋白肽生物素-IKVAV转移到改性表面上。作为生物测定，我们将LRM 55星形胶质瘤和原代大鼠海马神经元接种在图案化水凝胶上。我们发现这两种细胞类型选择性地粘附到生物素结合蛋白质图案化的区域。荧光和明场模式的显微镜被用来评估细胞附着和细胞形态的改性表面。发现LRM 55细胞仅附着于水凝胶的蛋白质压印区域。神经元表现出显着的神经突起延伸后，72它在体外，并保持活力的蛋白质冲压领域超过4周。图案化的神经元发展功能活跃的突触，如通过摄取染料FM 1 - 43 FX所测量的。这项研究的结果表明，水凝胶表面可以用多种蛋白质图案化，以指导细胞生长和附着。(C)2007 Elsevier B. V.保留所有权利。
Biochemical surface modification has been used to direct cell attachment and growth on a biocompatible gel surface. Acrylamide-based hydrogels were photo-polymerized in the presence of an acroyl-streptavidin monomer to create planar, functionalized surfaces capable of binding biotin-labelled proteins. Soft protein lithography (microcontact printing) of proteins was used to transfer the biotinylated extracellular matrix proteins, fibronectin and laminin, and the laminin peptide biotin-IKVAV, onto modified surfaces. As a biological assay, we plated LRM55 astroglioma and primary rat hippocampal neurons on patterned hydrogels. We found both cell types to selectively adhere to areas patterned with biotin-conjugated proteins. Fluorescence and bright-field modes of microscopy were used to assess cell attachment and cell morphology on modified surfaces. LRM55 cells were found to attach to protein-stamped regions of the hydrogel only. Neurons exhibited significant neurite extension after 72 It in vitro, and remained viable on protein-stamped areas for more than 4 weeks. Patterned neurons developed functionally active synapses, as measured by uptake of the dye FM1-43FX. Results from this study suggest that hydrogel Surfaces can be patterned with multiple proteins to direct cell growth and attachment. (C) 2007 Elsevier B.V. All rights reserved.