Function of ID1 protein in human cord blood-derived neural stem-like cells

The effect of dominant-negative regulator of basic helix-loop-helix (bHLH) transcription factors, an ID1 protein, on growth and differentiation of neural stem-like cell line derived from human umbilical cord blood (HUCB-NSC) was investigated. This nontransformed, mesodermal germ layer-originated line contains high levels of ID1 protein, whose intercellular distribution reflects HUCB-NSC differentiation status. Whereas cells remained undifferentiated and self-renewing in serum-free (SF) cultures, ID1 protein, although highly expressed, did not attain cell nuclei and was localized mainly in cytoplasm. In long-term-expanded cultures of partially committed (primed) HUCB-NSC grown in a low serum concentration (LS cultures) ID1 protein became translocated toward cell nuclei. Further neuronal differentiation of the cells, either spontaneous in the presence of serum or induced by neuromorphogens (dBcAMP, RA), resulted in almost complete depletion of ID1 mRNA and protein. Accordingly, HUCB-NSC transfectants overexpressing the ID1 gene were significantly inhibited in their differentiation. Notably, only neuronal and not glial development was affected after ID1 overexpression. A similar gain-of-function effect of ID1 transfection was observed in human NSC-like line (DEV) of medullobastoma origin, which is constitutively devoid of ID1 expression. Thus, our results on HUCB-NSC confirm further its neural-specific behavior and the crucial role of ID1 protein as a potent negative regulator of neural stem cell differentiation, pointing out that this protein distribution between cytoplasmic and nuclear cell compartments can be one of the most important steps in differentiation signal transduction.