A novel indirect co-culture model of primary neurons and glia

Direct and indirect co-culture systems provide feasible solutions for studying multicellular interactions in vitro, with the latter serving as an established paradigm for dissecting contact-independent cellular regulation. Current indirect co-culture systems primarily include traditional Transwell and the recently developed architecture utilizing paraffin spheres to compartmentalize cells cultured on distinct substrates. However, these models may not always be suitable.

We established a novel indirect co-culture model by using a soldering iron to fabricate protruding structures on culture plates, thereby separating cells grown on different substrates.

This model has been successfully applied to investigate multiple aspects of neuron-glia interactions, including microglial soluble factor-mediated neurotrophic support, neurotoxicity of pro-inflammatory microglia, and lipid shuttling between neurons and glial cells.

Our model offers several distinct advantages: streamlined operation, elimination of exogenous separator contamination, on-demand cell density control, and simplified sampling procedures, thereby enhancing experimental reproducibility and physiological relevance.

The novel indirect co-culture model expands the available platform choices, enabling researchers to select optimized configurations based on specific experimental requirements.