Protocol to generate PDMS topographical patterns for hiPSC-derived or rat primary neuronal cultures
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Updated: Sep 13, 2025
Stencil Micropatterning of Human Pluripotent Stem Cells for Probing Spatial Organization of Differentiation Fates
Published on: June 17, 2016
Anna-Christina Haeb1, Mireia Olives-Verger2, Jordi Soriano2
1Laboratory 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.
Abstract:
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.1.
