Integrated functional neuronal network analysis of 3D silk-collagen scaffold-based mouse cortical culture

• Published in: STAR protocols Vol. 2; no. 1; p. 100292
• Main Authors: Dingle, Yu-Ting L., Bonzanni, Mattia, Liaudanskaya, Volha, Nieland, Thomas J.F., Kaplan, David L.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 19-03-2021; Elsevier
• Subjects: Animals; Cell culture; Cell Culture Techniques; Cell Differentiation; Cerebral Cortex - cytology; Cerebral Cortex - metabolism; Collagen - chemistry; Mice; Microscopy; Nerve Net - cytology; Nerve Net - metabolism; Neural Networks, Computer; Neurons - cytology; Neurons - metabolism; Neuroscience; Protocol; Silk - chemistry; Tissue engineering; Tissue Scaffolds - chemistry; Cell culture; Neuroscience; Microscopy; Tissue engineering
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2020.100292

Bioengineered 3D tunable neuronal constructs are a versatile platform for studying neuronal network functions, offering numerous advantages over existing technologies and providing for the discovery of new biological insights. Functional neural networks can be evaluated using calcium imaging and quantitatively described using network science. This protocol includes instructions for fabricating protein-based composite scaffolds, 3D in vitro culture of embryonic mouse cortical neurons, virally induced expression of GCaMP6f, wide-field calcium imaging, and computational analysis with open-source software and custom MATLAB code.
For complete details on the use and execution of this protocol, please refer to Dingle et al. (2020).
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•Customizable 3D scaffold system to recapitulate in vitro neuronal networks•Optical mesoscale detection of neuronal activity•Structural, molecular, and functional characterization of the 3D in vitro system•Network science approach reveals millimeter-scale topologies
Bioengineered 3D tunable neuronal constructs are a versatile platform for studying neuronal network functions, offering numerous advantages over existing technologies and providing for the discovery of new biological insights. Functional neural networks can be evaluated using calcium imaging and quantitatively described using network science. This protocol includes instructions for fabricating protein-based composite scaffolds, 3D in vitro culture of embryonic mouse cortical neurons, virally induced expression of GCaMP6f, wide-field calcium imaging, and computational analysis with open-source software and custom MATLAB code.