I'm growing P0-P3 mouse hippocampus neurons and noticed the development of these clustered cells around DIV2-3. The culture is now about 12 days. What are these cells and how do I prevent future growth?
Protocol: Papain x 8-10 minutes then triturate with Media (NBM/L-glutamine/B27). I will say that I trialed FBS to my trituration media this time but there is no FBS in culture media.
Hi! It’s hard to know for sure without IHC or any objective way to mark them. Is it the first time you’re following this protocol? What have you gotten in your previous attempts/ what were you expecting?
From my experience in neurodifferention, these look like neuronal rosettes, which also agrees with their appearance timing. This could be due to poor initial cell dissociation or just the medium/substrate you’re using. It’s not necessarily a bad thing, even your objective is a neuronal monolayer, you could just dissociate and re-plate them in neuronal conditions.
Thank you for your response! 'm attempting to grow hippocampus in culture from post-natal mice. It's my first time adding FBS to the media of an old protocol. Otherwise, everything is the same. Ive attached a picture of previous attempts.
How would I go about dissociating and repeating?
I don't know if they were initially free-floating..they certainly are not now. Yes coated with PLL+laminin
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