Differentiation of neurons from neural precursors generated in floating spheres from embryonic stem cells
1 Department of Otolarygology, EENT Hospital of Shanghai Medical College of Fudan University, Institute of Biomedical Sciences, Fudan University Shanghai, 200031, PR China
2 Departments of Otolaryngology-HNS and Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA
3 Department of Otology and Laryngology, Harvard Medical School, Boston MA 02115, USA
4 Eaton-Peabody Laboratory. Massachusetts Eye and Ear Infirmary, Boston, MA 02114, USA
5 Program in Speech and Hearing Bioscience and Technology, Division of Health Science and Technology, Harvard and MIT, Cambridge, MA 02139, USA
6 Department of Neuroscience, University of Virginia, Charlottesville, VA 22908, USA
7 Department of Otolaryngology, University of Virginia, Charlottesville, VA 22908, USA
BMC Neuroscience 2009, 10:122 doi:10.1186/1471-2202-10-122Published: 24 September 2009
Neural differentiation of embryonic stem (ES) cells is usually achieved by induction of ectoderm in embryoid bodies followed by the enrichment of neuronal progenitors using a variety of factors. Obtaining reproducible percentages of neural cells is difficult and the methods are time consuming.
Neural progenitors were produced from murine ES cells by a combination of nonadherent conditions and serum starvation. Conversion to neural progenitors was accompanied by downregulation of Oct4 and NANOG and increased expression of nestin. ES cells containing a GFP gene under the control of the Sox1 regulatory regions became fluorescent upon differentiation to neural progenitors, and ES cells with a tau-GFP fusion protein became fluorescent upon further differentiation to neurons. Neurons produced from these cells upregulated mature neuronal markers, or differentiated to glial and oligodendrocyte fates. The neurons gave rise to action potentials that could be recorded after application of fixed currents.
Neural progenitors were produced from murine ES cells by a novel method that induced neuroectoderm cells by a combination of nonadherent conditions and serum starvation, in contrast to the embryoid body method in which neuroectoderm cells must be selected after formation of all three germ layers.