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

STAR Protoc. 2021 Jan 25;2(1):100292. doi: 10.1016/j.xpro.2020.100292. eCollection 2021 Mar 19.

Abstract

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).

Keywords: Cell culture; Microscopy; Neuroscience; Tissue engineering.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Differentiation
  • Cerebral Cortex / cytology
  • Cerebral Cortex / metabolism*
  • Collagen / chemistry*
  • Mice
  • Nerve Net / cytology
  • Nerve Net / metabolism*
  • Neural Networks, Computer*
  • Neurons / cytology
  • Neurons / metabolism*
  • Silk / chemistry*
  • Tissue Scaffolds / chemistry*

Substances

  • Silk
  • Collagen