Basic Cell Culture Protocols

Volume 946 of the series Methods in Molecular Biology pp 479-506


Methods to Culture, Differentiate, and Characterize Neural Stem Cells from the Adult and Embryonic Mouse Central Nervous System

  • Sharon A. LouisAffiliated withSTEMCELL Technologies Email author 
  • , Carmen K. H. MakAffiliated withSTEMCELL Technologies
  • , Brent A. ReynoldsAffiliated withDepartment of Neurosurgery, University of Florida

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Since the discovery of neural stem cells (NSC) in the embryonic and adult mammalian central nervous system (CNS), there have been a growing numbers of tissue culture media and protocols to study and functionally characterize NSCs and its progeny in vitro. One of these culture systems introduced in 1992 is referred to as the Neurosphere Assay, and it has been widely used to isolate, expand, differentiate and even quantify NSC populations. Several years later because its application as a quantitative in vitro assay for measuring NSC frequency was limited, a new single-step semisolid based assay, the Neural Colony Forming Cell (NCFC) assay was developed to accurately measure NSC numbers. The NCFC assay allows the discrimination between NSCs and progenitors by the size of colonies they produce (i.e., their proliferative potential). The evolution and continued improvements made to these tissue culture tools will facilitate further advances in the promising application of NSCs for therapeutic use.

Key words

Murine Embryonic neural stem cells Subventricular zone Neurospheres Differentiation CNS Culture Stem cells