how to passage neurospheres? - (Aug/05/2008 )

hi there
I would like to ask about how to passage neurospheres in right way? i have tried use mechanically dissociation and trypsinize method, but my neurospheres still in undissociated and yet will die.
please help me!
thanks....

I have not done it for neurospheres but know for mammospheres. We do trypsinization for ~3 min (spheres in 1ml TE) followed by gentle pipetting to get most single cells.

Use Accutase, incubate for a few minutes and triturate gently to get single cell suspension