Dear colleagues, I'm planning to start neuron mouse co-culture with astrocyte and microglia but I heard that it's difficult to keep it up. Any advices?
possibly you could make brain slice cultures - then all cell types are there, they can be maintained for a month easily (so long as you maintain sterility and don't keep them in medium with antibiotics). Microglia are well-ramified and surveying from about div9 until ~div24, astrocytes and neurons are also quite 'normal' we published a CSHL protocol and there are also good protocols published by others.
Hi Daniel Kalinowski
Primary mouse microglia cultures from P2 pups retain their characteristic morphology and function after a few passages (around 3-5) whereas astrocytes are more robust and can be passaged more frequently (around 8-10 passages). I do not have much experience in primary neuronal culture from P2 pups.
Thanks,
The problem is that I want to make up triculture from mouse brain in one Petri dish. I heard that glia cells are so moody, moreover I couldn't find any good protocol for mouse, many of them involved rat though.
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