gms | German Medical Science

56. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)
3èmes journées françaises de Neurochirurgie (SFNC)

Deutsche Gesellschaft für Neurochirurgie e. V.
Société Française de Neurochirurgie

07. bis 11.05.2005, Strasbourg

Epigenetic induction of phenotype-specific differentiation in long-term propagated human fetal-derived neural precursor cell cultures

Epigenetische Induktion von Phänotyp-spezifischer Differenzierung expandierter humaner fötalen neuronalen Vorläuferzellen

Meeting Abstract

Suche in Medline nach

  • corresponding author J. Maciaczyk - Labor für Molekulare Neurochirurgie, Abt. für Funktionelle und Stereotaktische Neurochirurgie, Universitätsklinikum Freiburg
  • D. Maciaczyk - Labor für Molekulare Neurochirurgie, Abt. für Funktionelle und Stereotaktische Neurochirurgie, Universitätsklinikum Freiburg
  • G. Nikkhah - Labor für Molekulare Neurochirurgie, Abt. für Funktionelle und Stereotaktische Neurochirurgie, Universitätsklinikum Freiburg

Deutsche Gesellschaft für Neurochirurgie. Société Française de Neurochirurgie. 56. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC), 3èmes journées françaises de Neurochirurgie (SFNC). Strasbourg, 07.-11.05.2005. Düsseldorf, Köln: German Medical Science; 2005. DocP208

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/dgnc2005/05dgnc0476.shtml

Veröffentlicht: 4. Mai 2005

© 2005 Maciaczyk et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

In vitro propagation of fetal tissue could serve as a source of cells for neural grafting in a clinical setting. However, the results of spontaneous differentiation of long-term propagated human fetal-derived neural precursor cells (hFNPCs) showed limited neurogenesis and a restricted phenotypic specification after multiple passages in vitro, and therefore led to the search for factors that could promote the generation of new neurons and efficient acquisition of different phenotypes of interest.

Cortex (CTX) ganglionic eminences (STR), ventral midbrains (VM) and spinal cord (SC) obtained from elective abortions during the first trimester of pregnancy were expanded as free floating neurospheres for a prolonged period in mitogen-containing medium. Cultures were then differentiated on the adhesive substrate in lowered oxygen concentration and further supplementation with the neurotrophins and growth factors (NT3, PDGF, CNTF, FCS, triiodothyronine). Moreover, the influence of numerous combinations of proteins involved in a phenotypic specification of the midbrain dopaminergic neurons during development of the CNS (Shh, FGF8, BDNF) on the induction of the tyrosine hydroxylase (TH) immunoreactivity was investigated. The acquisition of the specific phenotypes was revealed by immunostaining against type III β tubulin, MAP2, GFAP, Gal-C, neuroepithelial precursor cell marker - nestin and neuronal markers (TH, GABA).

Using a new differentiation protocol, we found an overall dramatic increase of MAP2 positive neurons (50% of total cells) in NT3 supplemented cultures and GFAP expressing glia (up to 70% of total cells) in cultures exposed to FCS/CNTF. Moreover, BDNF but not Shh and FGF8 resulted in a significant increase of TH expressing cells (15% of neurons) after long-term proliferation and serial passaging of neurospheres derived from the fetal VM and CTX.

The present study demonstrates the successful long-term proliferation of hFNPCs that retain the responsiveness to epigenetic signals after prolonged propagation. These cultures under appropriate serum-free culture conditions and without genetic manipulation, are capable of generating large amounts of neurons or glia and specific neurotransmitter phenotypes. Thus, long-term expanded hNPCs may be a realistic and large-scale cell source for neural grafting in patients with neurodegenerative diseases.