Efficient production of neural stem cells and neurons from embryonic stem cells.

We have developed a simple method to efficiently produce a large number of neural stem cells and neurons from mouse embryonic stem (ES) cells. When cultured in astrocyte-conditioned medium (ACM) with mitogens (FGF-2 and EGF) under free-floating conditions, colonies of undifferentiated ES cells give rise to neural stem spheres (NSSs), composed of plentiful neural stem cells. Subsequent culture of the NSSs on an adhesive substrate with mitogens results in the migration of neural stem cells onto the substrate. These cells can be expanded, preserved by freezing, and differentiated into functional neurons. Neural stem cells and neurons provided by this NSS method may be valuable as potential donor cells for neuronal transplantation and also as convenient alternatives to tissue-derived neural cells. Copyright 2004 Lippincott Williams & Wilkins