Welle Alexander, Horn Siegfried, Schimmelpfeng Jutta, Kalka Dorothee
Institute for Medical Engineering and Biophysics, Forschungszentrum Karlsruhe, Karlsruhe, Germany.
J Neurosci Methods. 2005 Mar 30;142(2):243-50. doi: 10.1016/j.jneumeth.2004.08.011.
The in vitro assembling of cellular networks offering control over cell positions and connectivities by patterned culture substrates is a valuable tool for neuroscience research and other applications in cell biology. We developed a versatile technique based on polymer surface modification which allows the patterning of different cell lines for advanced tissue engineering, among them are Pheochromocytoma cells (PC-12). In contrast to other techniques applied for surface patterning, the presented photo patterning by deep UV irradiation is applicable to the widely used cell culture substrate material polystyrene (PS) and should be easily performed in most laboratories. Irradiation of polystyrene with UV radiation of lambda = 185 nm yields mainly carboxyl groups at the polymer surface which can be used to control the spontaneous competitive protein adsorption from serum containing culture media [Welle A, Gottwald E. UV-based patterning of polymeric substrates for cell culture applications. Biomed. Microdev. 2002;4:33-41] or to serve as defined coupling sites for controlled protein/peptide immobilization. Extending our previous studies on patterning hepatoma cells and fibroblasts via spatially defined plasma protein adsorption, we here describe an advanced application to produce patterns of cell repellent albumin domains and cell attractive laminin regions for the patterning of Pheochromocytoma cells.
通过图案化培养底物来体外组装细胞网络,从而控制细胞位置和连接性,这对于神经科学研究以及细胞生物学的其他应用而言是一种有价值的工具。我们基于聚合物表面改性开发了一种通用技术,该技术可对不同细胞系进行图案化,以用于先进的组织工程,其中包括嗜铬细胞瘤细胞(PC - 12)。与用于表面图案化的其他技术不同,本文所介绍的通过深紫外辐射进行的光图案化适用于广泛使用的细胞培养底物材料聚苯乙烯(PS),并且在大多数实验室中应该都能轻松完成。用波长λ = 185 nm的紫外辐射照射聚苯乙烯,主要会在聚合物表面产生羧基,这些羧基可用于控制含血清培养基中蛋白质的自发竞争性吸附[韦勒A,戈特瓦尔德E。用于细胞培养应用的聚合物底物的紫外基图案化。生物医学微器件。2002;4:33 - 41],或者用作可控蛋白质/肽固定的特定偶联位点。在我们之前关于通过空间定义的血浆蛋白吸附对肝癌细胞和成纤维细胞进行图案化的研究基础上,我们在此描述一种先进应用,即生成细胞排斥性白蛋白结构域和细胞吸引性层粘连蛋白区域的图案,用于嗜铬细胞瘤细胞的图案化。