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Published on: June 17, 2016
Protocol to generate PDMS topographical patterns for hiPSC-derived or rat primary neuronal cultures
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.
This study details a protocol for creating patterned polydimethylsiloxane (PDMS) substrates using soft lithography. These substrates enable the in vitro culturing of neuronal networks with defined connectivity for research.
Area of Science:
- Neuroscience
- Biomaterials Science
- Microfabrication
Background:
- Soft lithography is a key technique for fabricating custom substrates.
- Polydimethylsiloxane (PDMS) is a versatile material for microfluidic and biomaterial applications.
- Controlling neuronal network architecture is crucial for understanding neural function.
Purpose of the Study:
- To present a detailed protocol for generating topographical patterns on PDMS using soft lithography.
- To establish a method for culturing human induced pluripotent stem cell (hiPSC)-derived and rat neurons on these patterned substrates.
- To enable the creation of neuronal networks with imprinted connectivity for in vitro studies.
Main Methods:
- Development of a two-level resin master mold utilizing soft lithography.
- Fabrication of topographical PDMS casts from the master mold.
- Culturing of hiPSC-derived and rat neurons on patterned PDMS substrates.
- Data acquisition using calcium imaging.
Main Results:
- Successful generation of PDMS topographical patterns.
- Demonstration of neuronal network formation on patterned PDMS.
- Imprinted connectivity within cultured neuronal networks.
- Feasibility of calcium imaging for data acquisition.
Conclusions:
- The protocol provides a reliable method for fabricating patterned PDMS substrates for neuronal cultures.
- This technique allows for the precise control of neuronal network architecture in vitro.
- The developed method facilitates the study of neuronal connectivity and function.
