Method for neural stem cell differentiation
Reference Number TO 0017
Neural stem cells have been isolated from various regions of the adult mammalian brain. These cells can be differentiated in vivo and in vitro into neurons and glia, which is a useful approach for tissue regeneration or pharmaceutical vehicle.
A variety of methods have been described to differentiate neural stem cells (Björklund & Lindvall, Nat Neurosci, 3, 537 (2000); Björklund & Lindvall, Nature, 405, 892 (2000); Cameron et al., J Neurobiol, 36, 287 (1998); McKay, Nature, 406, 361 (2000); Wachs et al., Lab Invest. 83: 949 (2003)), but all of these have the following disadvantages:
· They are not well suited for high-throughput technologies necessary
for pharmacotechnological use.
· They often rely on embryonic stem cells, bearing ethical problems
and legal regulations.
· The protocols for in vitro differentiation involve multiple and
· The yield of differentiated neurons is low.
· The monitoring of successful differentiation is difficult and
The invention presented here aimed to overcome these disadvantages and is characterized by:
· It is suited for high-throughput cell biotechnology.
· It is developed for adult neural stem cells.
· It is a single step protocol.
· The yield of neurons is high.
· The monitoring of differentiation can be adapted to high-throughput
methods, e. g. such as FACS analysis.
Fig. 1: Neuronal morphology of a neuronal stem cell differentiated in vitro by inhibition of GSK3ß. Source: Martin H. Maurer.
The protocol of this invention is based on the culture of neural stem cells, expanded and differentiated by the inhibition of the Wnt signalling pathway, for example by pharmacological inhibition of its central enzyme, Glycogen Synthase Kinase 3ß (GSK3ß).
The differentiation protocol can be used to provide a large number of neurons in vitro which then can be transplanted into the brain to regenerate tissue lost due to neurodegeneration or trauma. It can be adapted to the large-scale production requirements of pharmaceutical biotechnology.
US, CA, and DE patent applications are pending.
Maurer et al. (2007), J. Proteome Res. 6, 1198-1208.