Foetal mouse neural stem cells give rise to ependymal cells in vitro

  • Mokry J
  • Karbanova J
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Abstract

NSCs are responsible for the generation of CNS cell types derived from
the neural tube. Published data resulting from experiments studying the
differentiation of NSCs in vitro or in vivo have confirmed their
spontaneous tripotency, i.e. their ability to generate cells of the
neuronal, astroglial and oligodendroglial lineages. The relationship
between NSCs generated in vitro and ependymal cells has not yet been
studied. To confirm that ependymal cells can also be produced by NSCs,
we utilized the neurosphere assay, which permits isolation and
cultivation of NSCs. Cells from the forebrain of E14-15 Balb/c foetuses
were grown in DMEM/F12-N2 medium supplemented with EGF and FGF-2 to form
multicellular neurospheres. After 3 to 8 passages, neurospheres were
plated on surfaces coated with poly-L-lysine, polyornithine and/or
laminin in dishes containing the same medium where cytokines were
replaced with serum. Under these conditions, neurosphere cells spread
over the surface forming a cellular layer consisting of beta-III
tubulin(+) neuronal, GFAP(+) astroglial and O4(+) oligodendroglial
cells. When these cells were cultivated for prolonged periods, they
formed islands of epitheloid cells. Following 2 to 3 weeks in vitro,
ependymal cells with beating cilia appeared among these cells. Ciliated
ependymal cells were observed in small clusters or as single cells
scattered in certain areas. Confocal microscopy confirmed the presence
of alpha-tubulin-immunoreactive cilia arranged in tufts located on the
apical surface of epitheloid cells. Our data indicate that ependymal
cells are spontaneously derived from NSCs.

Author-supplied keywords

  • neural stem cells; neurospheres; ependymal cells;

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Authors

  • J Mokry

  • J Karbanova

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