Hi guys!
I'm trying to co-culture neurons and astrocytes. I'm using a protocol that first grows the neurons into plates (let then grow) and then I put in the astrocytes (fully grown) and supplement the neurobasal with FBS (5% and 10% - currently trying two concentrations of it). However, in my last attempt, I noticed a detachment of neurons after the astrocytes... I don't know why... I am actually working with 3 concentrations of astrocytes (60, 50 and 40% of the neurons seeded in the plate) .
I don't have the time and resources to co-culture with films and whatever.... has anyone done this protocol or know how to do it? Thanks!!
Another question, Can I trypsinize neurons? I've seen some protocols that do backwards... so I don't know what to do.
Hi,
To answer your second question, if it is a primary neuronal culture, you can not trypsinize and replate them. You can scrape your neuronal cells and perform experiment such as western blot, QPCR etc after respective processing.However, with secondary neuronal cells, you can try trypsinization and replate them.
For coculture experiments, again, there are various protocols. If your intention is only to see the effect of factors released by astrocyte on neuronal culture, you can try adding conditioned medium from astrocyte culture in to your neuronal culture.
I hope this helps.
Best,
Alpana
Thanks, Alpana
I'm trying to grow they together, however in the protocol that I have, the neurons are seeded and then we put the astrocytes. If I use the astrocytes medium, all the neurons die. Because of it, I'm using neurobasal with FBS, this is the only way to keep both surviving. The problem is, when I put the astrocytes over the neurons, a lot of then detached and I dont know why! =/
Hi again,
Here is a link:
https://experiments.springernature.com/articles/10.1007/978-1-61779-452-0_22
This paper plates a feeder layer of astrocytes first and on top of that, adds the neuronal culture. Go through this protocol. Maybe this one will work for you.
All the best,
Alpana
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