Isolation of neural stem cells from neural tissues using the neurosphere technique

Curr Protoc Stem Cell Biol. 2010 Nov;Chapter 2:Unit2D.6. doi: 10.1002/9780470151808.sc02d06s15.


This unit describes protocols for the derivation, characterization, and expansion of neural stem cell (NSC) lines from the adult mouse subventricular zone (mNSCs), embryonic mouse brain and from the human fetal brain (hNSCs). NSCs can be isolated by enzymatic digestion of specific regions (NSCs niches) of the central nervous system (CNS) and grown in suspension. By using this methodology, NSCs form spherical clusters called neurospheres, which are mechanically dissociated to a single-cell suspension and replated in the selective culture medium. Removal of growth factors and plating cells on an adherent substrate allows cells to differentiate into neurons, astrocytes, and oligodendrocytes, the main cell type of the CNS. Correct culturing of NSCs, according to this methodology, will allow cells to expand over 100 passages without alteration of cell karyotype, growth ability, and differentiation potential.

MeSH terms

  • Animals
  • Biological Assay
  • Cell Aggregation
  • Cell Count
  • Cell Differentiation
  • Cell Separation / methods*
  • Cells, Cultured
  • Cerebral Ventricles / cytology
  • Clone Cells
  • Cryopreservation
  • Humans
  • Mice
  • Multipotent Stem Cells / cytology
  • Nerve Tissue / cytology*
  • Neural Stem Cells / cytology*