Lineage-restricted neural precursors can be isolated from both the mouse neural tube and cultured ES cells

We have previously identified multipotent neuroepithelial (NEP) stem cells and lineage-restricted, self-renewing precursor cells termed NRPs (neuron-r estricted precursors) and GRPs (glial-restricted precursors) present in the developing rat spinal cord (A. Kalyani, K. Hobson, and M. S. Rao, 1997, De v. Biol. 186, 202-223; M. S. Rao and M. Mayer-Proschel, 1997, Dev. Biol. 18 8, 48-63; M. Mayer-Proschel, A. J. Kalyani, T. Mujtaba, and M. S. Rao, 1997 , Neuron 19, 773-785). We now show that cells identical to rat NEPs, NRPs, and GRPs are present in mouse neural tubes and that immunoselection against cell surface markers E-NCAM and A2B5 can be used to isolate NRPs and GRPs, respectively. Restricted precursors similar to NRPs and GRPs can also be i solated from mouse embryonic stem cells (ES cells). ES cell-derived NRPs ar e E-NCAM immunoreactive, undergo self-renewal in defined medium, and differ entiate into multiple neuronal phenotypes in mass culture. ES cells also ge nerate A2B5-immunoreactive cells that are similar to E9 NEP-cell-derived GR Ps and can differentiate into oligodendrocytes and astrocytes. Thus, lineag e restricted precursors can be generated in vitro from cultured ES cells an d these restricted precursors resemble those derived from mouse neural tube s. These results demonstrate the utility of using ES cells as a source of l ate embryonic precursor cells. (C) 1999 Academic Press.