NMDA tox and Rat Cerebellar Granular Cells ! - NMDA tox assay for Rat CGC's ???? (Mar/02/2006 )

Hey Folks, I'm culturing day 8 rat cerebellar granular cells and trying to get a toxicity assay for NMDA going. It seems that every paper I pick up grows the cells in something different.

Currently I'm using Neurobasal A medium and B27 supplement + Mg+2

What is the optimal buffer/media to use for the assay? Can I use the neurobasal/B27 instead of a specific tox assay buffer?

Is exposure time to NMDA important is leaving it on for an hour ok?

Also there is a huge variety of days on which the CGC's are used, sometimes Day 10 sometimes Day 14. Any suggestions?

Thanks very much for your help, it's massively appreciated !!!!!!!!!!!!!!!!!!!!!!!

-pj263-

QUOTE (pj263 @ Mar 2 2006, 05:50 PM)

You can use Neurobasal/B27....but you have to keep cells at 9% oxygen. However, if you are doing you're experiments in ambient conditions then you cannot use Neurobasal instead you should use Hibernate A/B27. It is designed to keep the pH stable in ambient conditions.

NMDA exposure time....I usually do 24hrs. You could do a time course try 1hr, 6hr, 18hrs and 24 hrs.

Hope this helps

JP

-JP2877-

Thanks so much JP

I'm digging through so many papers on NMDA protecting neuronal survival as well as NMDA-induced tox in rat CGC's that my was feeling pretty scrambled.

cheers

Paul

-pj263-

QUOTE (pj263 @ Mar 21 2006, 03:25 PM)

Thanks so much JP

I'm digging through so many papers on NMDA protecting neuronal survival as well as NMDA-induced tox in rat CGC's that my was feeling pretty scrambled.

cheers

Paul

We tried neurobasal and the cells multilayered, and dont' even look like neurons, they look like fibroblasts, and assorted epithelial shapes...But I don't know the O2 percentage either. Very odd, but I don't culture neurons..zero experience

-viper-