Cell patterning on photolithographically defined parylene-C: SiO2 substrates.

Cell patterning on photolithographically defined parylene-C: SiO2 substrates.
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DOI:
10.3791/50929
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发表时间:
2014-03-07
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Murray AF
Murray AF
中科院分区:
其他
文献类型:
--
作者:
Hughes MA;Brennan PM;Bunting AS;Shipston MJ;Murray AF

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细胞模式平台支持广泛的研究目标,如预先确定的体外神经元网络的构建和细胞生理学的某些核心方面的探索。为了轻松地将细胞图谱与多电极阵列(MEAs)和硅基“芯片实验室”技术相结合,需要一种微加工兼容的协议。我们描述了一种利用聚合物聚苯乙烯- c沉积在SiO2晶片上的方法。光刻技术能够在微米级分辨率下精确可靠地制作聚苯乙烯- c的图案。随后通过浸泡在胎牛血清(或另一种特定的激活溶液)中的激活产生底物,其中培养的细胞分别粘附或被聚对二甲苯或二氧化硅区域排斥。该技术允许广泛的细胞类型(包括原代小鼠海马细胞、HEK 293细胞系、人类神经元样畸胎癌细胞系、原代小鼠小脑颗粒细胞和原代人类胶质瘤来源的干细胞样细胞)的模式化。然而有趣的是,这个平台并不是通用的;反映了细胞特异性粘附分子的重要性。这种细胞图像化工艺具有成本效益,可靠,重要的是可以纳入标准微加工(芯片制造)协议,为微电子技术的集成铺平了道路。
Cell patterning platforms support broad research goals, such as construction of predefined in vitro neuronal networks and the exploration of certain central aspects of cellular physiology. To easily combine cell patterning with Multi-Electrode Arrays (MEAs) and silicon-based ‘lab on a chip’ technologies, a microfabrication-compatible protocol is required. We describe a method that utilizes deposition of the polymer parylene-C on SiO2 wafers. Photolithography enables accurate and reliable patterning of parylene-C at micron-level resolution. Subsequent activation by immersion in fetal bovine serum (or another specific activation solution) results in a substrate in which cultured cells adhere to, or are repulsed by, parylene or SiO2 regions respectively. This technique has allowed patterning of a broad range of cell types (including primary murine hippocampal cells, HEK 293 cell line, human neuron-like teratocarcinoma cell line, primary murine cerebellar granule cells, and primary human glioma-derived stem-like cells). Interestingly, however, the platform is not universal; reflecting the importance of cell-specific adhesion molecules. This cell patterning process is cost effective, reliable, and importantly can be incorporated into standard microfabrication (chip manufacturing) protocols, paving the way for integration of microelectronic technology.
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