Protocol for co-culture of primary neurons and organotypic hippocampal brain slices from mice
Year: 2025
Type: article
Abstract: Here, we present a protocol for establishing a co-culture system combining organotypic hippocampal slices with primary mouse neurons to model neuronal interactions. We describe steps for preparing hippocampal slices by whole-brain dissection, tissue isolation, and sectioning. We then detail procedures for plating and culturing on membrane inserts, isolating and culturing primary neurons, and integrating to form the co-culture. The system is compatible with transgenic mice or genetically modified neurons, enabling investigation of molecular mechanisms and therapeutic targets. For complete details on the use and execution of this protocol, please refer to Zhou et al. 1 • Steps for preparation of organotypic hippocampal brain slices from adult mice • Instructions for plating and culturing hippocampal brain slices • Guidance on isolating and culturing primary mouse neurons • Procedures for co-culturing hippocampal brain slices with primary mouse neurons Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Here, we present a protocol for establishing a co-culture system combining organotypic hippocampal slices with primary mouse neurons to model neuronal interactions. We describe steps for preparing hippocampal slices by whole-brain dissection, tissue isolation, and sectioning. We then detail procedures for plating and culturing on membrane inserts, isolating and culturing primary neurons, and integrating to form the co-culture. The system is compatible with transgenic mice or genetically modified neurons, enabling investigation of molecular mechanisms and therapeutic targets.
Source: STAR Protocols
Authors: Linlin Zhou, Qihui Wu
Institutions: Tongji University, Shanghai East Hospital
Language: English
Subfield: Cellular and Molecular Neuroscience
Field: Neuroscience
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W4416619824