Hi all,
I am working with neural stem cells from SVZ in mice. Usually I get these cells from 4-7 days neonatal mice. this time I got cells from 11 days neonatal and result of my cell culture is completely different. All the primary plates show just clumps of cell.
my media is DMEM(F12)+B27+ gentamycine+ glutamin.
Dose anyone know how I can treat these clumps and gets neurospheres?
Hi Azadeh,
we use DMEM/F12+N2+P/S + EGF/FGF2 for neurospheres of adult SVZ and SGZ. SVZ usually does not make big problems in these conditions. Maybe they are growing more slowly than the early postnatal ones (we usually don't touch the cultures for 7-10 days before checking for spheres), and probably there is more tissue debris (myelin, dead cells, ...) in the initial cultures. Else might there be a problem with the tissue dissociation, which is probably different for the adult tissue. We use the Milteny adult brain dissociation kit, which works quite well.
I hope this helps.
Cheers,
Michael
Hi Azadeh,
we use DMEM/F12+N2+P/S + EGF/FGF2 for neurospheres of adult SVZ and SGZ. SVZ usually does not make big problems in these conditions. Maybe they are growing more slowly than the early postnatal ones (we usually don't touch the cultures for 7-10 days before checking for spheres), and probably there is more tissue debris (myelin, dead cells, ...) in the initial cultures. Else might there be a problem with the tissue dissociation, which is probably different for the adult tissue. We use the Milteny adult brain dissociation kit, which works quite well.
I hope this helps.
Cheers,
Michael
Thank you so much Michael for your help. I think problem can be with tissue dissociation. I used 26 pub mice and I think it was too much tissue for Dnase enzyme. Next time I will use less number.
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