Leukemia inhibitory factor favours neurogenic differentiation of long-term propagated human midbrain precursor cells.
There is a lot of excitement about the potential use of multipotent neural stem cells for the treatment of neurodegenerative diseases. However, the strategy is compromised by the general loss of multipotency and ability to generate neurons after long-term in vitro propagation. In the present study, human embryonic (5 weeks post-conception) ventral mesencephalic (VM) precursor cells were propagated as neural tissue-spheres (NTS) in epidermal growth factor (EGF; 20 ng/ml) and fibroblast growth factor 2 (FGF2; 20 ng/ml). After more than 325 days, the NTS were transferred to media containing either EGF+FGF2, EGF+FGF2+heparin or leukemia inhibitory factor (LIF; 10 ng/ml)+FGF2+heparin. Cultures were subsequently propagated for more than 180 days with NTS analyzed at various time-points. Our data show for the first time that human VM neural precursor cells can be long-term propagated as NTS in the presence of EGF and FGF2. A positive effect of heparin was found only after 150 days of treatment. After switching into different media, only NTS exposed to LIF contained numerous cells positive for markers of newly formed neurons. Besides of demonstrating the ability of human VM NTS to be long-term propagated, our study also suggests that LIF favours neurogenic differentiation of human VM precursor cells.
Andersen
Rikke K
RK
Department of Anatomy and Neurobiology, Institute of Medical Biology, University of Southern Denmark, Winsløwparken 21, DK-5000 Odense C, Denmark.
Widmer
Hans R
HR
Zimmer
Jens
J
Wahlberg
Lars U
LU
Meyer
Morten
M
D002454:Cell Differentiation; D002478:Cells, Cultured; D003470:Culture Media; D004815:Epidermal Growth Factor / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D008636:Mesencephalon / Q000166:cytology*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects; D013997:Time Factors
D003470:Culture Media; D053605:Leukemia Inhibitory Factor; D016222:Fibroblast Growth Factor 2; D004815:Epidermal Growth Factor; D006493:Heparin