Cryopreservation of Neurospheres

Tips for successful cryopreservation of primary neural stem and progenitor cells as neurospheres:

  • Freeze neurospheres at an early passage number to preserve the potential of the neural stem and progenitor cells.
  • Harvest neurospheres before they grow too large (100-200 μm). If neurospheres are too big (>200 μm), their survival after cryopreservation will be negatively affected.
  • Freeze whole neurospheres, do not break them up by mechanical dissociation, as this will increase the number of dead cells, significantly lowering the viability of the thawed culture.
  • Freeze intact neurospheres in the appropriate species-specific Complete NeuroCult™ Proliferation Medium (i.e. supplemented with rh EGF, rh bFGF and Heparin as appropriate) containing 10% DMSO (v/v), using controlled rate freezing containers.
For a detailed protocol, please refer to our TECHNICAL BULLETIN: Cryopreserving Neurospheres.

Please contact us at techsupport@stemcell.com with any questions about this procedure, or for more information on culturing neural stem & progenitor cells in our NeuroCult™ media.