NEUROEPITHELIAL STEM-CELLS FROM THE EMBRYONIC SPINAL-CORD - ISOLATION, CHARACTERIZATION, AND CLONAL ANALYSIS

Adherent cultures of E10.5 rat neuroepithelial cells (NEP cells) from the caudal neural tube require FGF (fibroblast growth factor) and CEE (chick embryo extract) to proliferate and maintain an undifferentiated phenotype in culture. Epidermal growth factor (EGF) does not support E10.5 NEP cells in adherent culture and NEP cells do not form EGF-depe ndent neurospheres. NEP cells, however, can be grown as FGF-dependent neurospheres. NEP cells express nestin and lack all lineage-specific m arkers for neuronal and glial sublineages, retain their pleuripotent c haracter over multiple passages, and can differentiate into neurons, a strocytes, and oligodendrocytes when plated on laminin in the absence of CEE. In clonal culture, NEP cells undergo self-renewal and generate colonies that vary in size from single cells to several thousand cell s. With the exception of a few single-cell clones, all other NEP-deriv ed clones contain more than one identified phenotype, with over 40% of the colonies containing A2B5, beta-111 tubulin, and GFAP-immunoreacti ve cells. Thus, NEP cells are multipotent and capable of generating mu ltiple neural derivatives. NEP cells also differentiate into motoneuro ns immunoreactive for choline acetyl transferase (ChAT) and the low-af finity neurotrophin receptor (p75) in both mass and clonal culture. Do uble labeling of clones for ChAT and glial, neuronal, or oligodendrocy tic lineage markers shows that motoneurons always arose in mixed cultu res with other differentiated cells. Thus, NEP cells represent a commo n progenitor for motoneurons and other spinal cord cells. The relation ship of NEP cells with other neural stem cells is discussed. (C) 1997 Academic Press.