Hi there, I'm isolating neurons from PND1 guinea pigs and have noticed a bit of debris in the early days (this is day 4 plated) and a lot of variability between plates. Current protocol is dissociate with 50uL Trypsin in 5mL HBSS (wondering if I should add DNase?), spin and wash pellet and plate at 60x10^4 cells/well in a 24 well plate in Neurobasal/B27/L-glut/PenStrp/HEPES/10% horse serum. Leave them overnight then change to a serum free media. I've attached a photo, any advice is appreciated
I don' see any debris actually. What I see are clumps of cells. I don't know, are you sure these are debris and not just clumps of cells? Make sure.
Judging from the picture, this should be okay to work with.
Are you sure it's not just your horse serum? I find horse serum to be quite 'bitty' on occasion. It doesn't affect anything, but you can always filter your serum (or the final media) before use.
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