Enhanced dopaminergic differentiation of human neural stem cells by synergistic effect of Bcl-xL and reduced oxygen tension |
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| Authors: | Christina Krabbe&dagger ,Elise Courtois&Dagger ,Pia Jensen,Jesper R. Jø rgensen&dagger ,Jens Zimmer,Alberto Martí nez-Serrano&Dagger , Morten Meyer |
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| Affiliation: | Department of Anatomy and Neurobiology, Institute of Medical Biology, University of Southern Denmark, Odense C, Denmark;
NsGene A/S, Ballerup, Denmark;
Department of Molecular Biology and Center of Molecular Biology Severo Ochoa, U.A.M.-C.S.I.C. Campus Cantoblanco, Madrid, Spain |
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| Abstract: | Neural stem cells constitute a promising source of cells for transplantation in Parkinson's disease, but a protocol for controlled dopaminergic differentiation is not yet available. Here we investigated the effect of the anti-apoptotic protein Bcl-xL and oxygen tension on dopaminergic differentiation and survival of a human ventral mesencephalic stem cell line (hVM1). hVM1 cells and a Bcl-xL over-expressing subline (hVMbcl-xL) were differentiated by sequential treatment with fibroblast growth factor-8, forskolin, sonic hedgehog, and glial cell line-derived neurotrophic factor. After 10 days at 20% oxygen, hVMbcl-xL cultures contained proportionally more tyrosine hydroxylase(TH)-positive cells than hVM1 control cultures. This difference was significantly potentiated from 11 ± 0.8% to 17.2 ± 0.2% of total cells when the oxygen tension was lowered to 3%. Immunocytochemistry and Q-PCR-analysis revealed expression of several dopaminergic markers besides of TH just as dopamine was detected in the culture medium by HPLC analysis. Although Bcl-xL-over-expression reduced cell death in the cultures, it did not alter the relative content of GABAergic, neurons, while the content of astroglial cells was reduced in hVMbcl-xL cell cultures compared with control. We conclude that Bcl-xL and lowered oxygen tension act in concert to enhance dopaminergic differentiation and survival of human neural stem cells. |
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| Keywords: | dopamine Parkinson's disease precursor cells tyrosine hydroxylase ventral mesencephalon |
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