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Cerebellar Granular Cells and NMDA tox
Posted by: pj263 (IP Hidden, New member, 2)
Date: March 8, 2006 12:51PM
Hi Folks, I'm trying to establish a protocol for culturing rat (day 9 post-natal) cerebellar granular cells to be used in NMDA toxicity assays but every paper I read has a different protocol.
If anyone has a protocol that they are having terrific success with, I'd love it if you could share it. There is so much variability as to when cells are used, what media, what buffer the Tox assay is performed in, how long the cells are exposed to NMDA etc etc, any help would be massively appreciated !!!!!!!!!!!!!!! Thank you so much
Re: Cerebellar Granular Cells and NMDA tox
Posted by: pj263 (IP Hidden, New member, 2)
Date: April 25, 2006 09:32AM
I had a very helpful response to my first request with respect to NMDA tox and rat cerebellar granular cells.
Could people please comment on the most effective buffer to use for the toxicity assay? Issues I'm concerned about are the amount of KCl in the tox buffer, presence of Mg+2, and the general media used. I'm confused as to why some assays require glycine to block the impact of Mg+2 while others don't. Some studies use Locke's Buffer. The cell density in 24 and 48 well culture plates seems to be hugely variable too. Comments on any of these parameters would be a fantastic help. Best wishes Paul
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