Hi! Could you tell me more about your model? From the text, it seems like you culture astrocytes and neurons separately and then mix them together. If you're using primary mouse cells, why not plate them together from the start? Apologies if I misunderstood something.

Alina Chaplygina Hi Alina! Thanks for your reply. In my model, cells were separately cultured and mixed as you said. I read some paper about tripartite synapses and most of them seems to use the model that I used. Now I'm trying other models too (as you recommended), but I'm little curious what models are the best.

Do you recommend to plate cells together? Does it generate synapses properly?

I think that co-culturing cells from the start is more physiologically relevant. Yes, when neurons and glia are plated together, they tend to form mature, functional synapses.

There can be some challenges with imaging, though, primary mixed cultures grow in layers and are often intertwined, so it might be worth experimenting with seeding density and surface area. If the density is too high, the cells will grow well, but imaging will be difficult, if it’s too low, they may not form proper synaptic connections.

Hi there,

• Neurons often detach when astrocytes are added too suddenly — precondition them by gradually mixing astrocyte-conditioned medium before direct co-culture.
• Seeding astrocytes first as a stable layer, then adding neurons, can reduce stress compared to adding astrocytes later.
• PDL coatings can degrade; adding laminin or refreshing coatings helps maintain neuron adhesion.
• For stronger, long-term neuron–astrocyte co-cultures, DendroTEK dPGA | Protease-Resistant Culture Matrix provides more stable adhesion than standard coatings.

Source: I work at DendroTEK Biosciences. www.dendrotek.ca