Enhanced dopaminergic differentiation of human neural stem cells by synergistic effect of Bcl-xL and reduced oxygen tension

Christina Krabbe, Elise Courtois, Pia Jensen, Jesper Jørgensen, Jens Zimmer, Alberto Martínez-Serrano, Morten Meyer

Research output: Contribution to journalJournal articleResearchpeer-review

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-x(L) and oxygen tension on dopaminergic differentiation and survival of a human ventral mesencephalic stem cell line (hVM1). hVM1 cells and a Bcl-x(L) over-expressing subline (hVMbcl-x(L)) 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-x(L) 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-x(L)-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-x(L) cell cultures compared with control. We conclude that Bcl-x(L) and lowered oxygen tension act in concert to enhance dopaminergic differentiation and survival of human neural stem cells.
Original languageEnglish
JournalJournal of Neurochemistry
Volume110
Issue number6
Pages (from-to)1908-20
Number of pages13
ISSN0022-3042
DOIs
Publication statusPublished - 2009

Keywords

  • Apoptosis Inducing Factor
  • Bromodeoxyuridine
  • Carbonyl Cyanide m-Chlorophenyl Hydrazone
  • Cell Count
  • Cell Differentiation
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • Dibutyryl Cyclic GMP
  • Dopamine
  • Embryonic Stem Cells
  • Fetus
  • Forskolin
  • Gene Expression
  • Green Fluorescent Proteins
  • HN Protein
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Ionophores
  • L-Lactate Dehydrogenase
  • Mesencephalon
  • Microtubule-Associated Proteins
  • Neurons
  • Oxygen
  • Phorbol Esters
  • Transfection
  • Tyrosine 3-Monooxygenase
  • bcl-X Protein

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