用于生成人诱导多能干细胞衍生或大鼠原代神经元培养物的聚二甲基硅氧烷（PDMS）拓扑图案的方案。
Protocol to generate PDMS topographical patterns for hiPSC-derived or rat primary neuronal cultures.
作者信息
Haeb Anna-Christina, Olives-Verger Mireia, Soriano Jordi
机构信息
Laboratory of Neuronal Stem Cells and Cerebral Damage, Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Institute of Neurosciences, University of Barcelona, 08036 Barcelona, Spain; Department of Condensed Matter Physics, Faculty of Physics, University of Barcelona, 08028 Barcelona, Spain; Universitat de Barcelona Institute of Complex Systems (UBICS), 08028 Barcelona, Spain.
Department of Condensed Matter Physics, Faculty of Physics, University of Barcelona, 08028 Barcelona, Spain; Universitat de Barcelona Institute of Complex Systems (UBICS), 08028 Barcelona, Spain.
出版信息
STAR Protoc. 2025 Aug 1;6(3):104010. doi: 10.1016/j.xpro.2025.104010.
Soft lithography is a promising technique to fabricate tailored substrates on polydimethylsiloxane (PDMS) casts. Here, we present a protocol to generate PDMS topographical patterns for neuronal networks in vitro. The protocol outlines the development of a two-level resin master mold with soft lithography and the creation of a topographical PDMS cast from the mold. We then detail the procedures for culturing either human induced pluripotent stem cell (hiPSC)-derived or rat neurons on the PDMS cast with topographical patterns and for acquiring data using calcium imaging. This protocol provides neuronal networks with imprinted connectivity. For complete details on the use and execution of this protocol, please refer to Montalà-Flaquer et al..
软光刻是一种在聚二甲基硅氧烷（PDMS）铸模上制造定制基板的有前途的技术。在此，我们展示了一种在体外为神经网络生成PDMS拓扑图案的方案。该方案概述了通过软光刻开发两级树脂母模以及从该模具创建拓扑PDMS铸模的过程。然后，我们详细介绍了在具有拓扑图案的PDMS铸模上培养人诱导多能干细胞（hiPSC）衍生的神经元或大鼠神经元以及使用钙成像获取数据的程序。该方案为神经网络提供了印记连接性。有关该方案的使用和执行的完整详细信息，请参考蒙塔拉 - 弗拉克尔等人的研究。
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