Fol. Biol. 2006, 52, 149-155
Foetal Mouse Neural Stem Cells Give Rise to Ependymal Cells in Vitro
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 β β-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 α α-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.
Keywords
neural stem cells, neurospheres, ependymal cells, ependymogenesis, in vitro, differentiation potential, mouse.
Funding
The work was supported by grant No. MSM 0021620820 from the Ministry of Education, Youth and Sports of the Czech Republic.
References
Copyright
This is an open-access article distributed under the terms of the Creative Commons Attribution License.