Induction of dopaminergic neurons from growth factor expanded neural stem/progenitor cell cultures derived from human first trimester forebrain

• Published in: Brain research bulletin 2006, Vol.70 (4), p.457-466
• Main Authors: Christophersen, Nicolaj S., Meijer, Xia, Jørgensen, Jesper R., Englund, Ulrica, Grønborg, Mette, Seiger, Åke, Brundin, Patrik, Wahlberg, Lars U.
• Format: Article
• Language: English
• Subjects: Animals; Basic Medicine; Biomedical Sciences; Biomedicinsk vetenskap; Brain Injuries - chemically induced; Brain Injuries - metabolism; Brain Injuries - pathology; Brain Injuries - surgery; Cell Count - methods; Cell Differentiation - drug effects; Cell Transplantation - methods; Cells, Cultured; Differentiation method; Dopamine - metabolism; Dopaminergic; Female; Fetus; Gene Expression Regulation - drug effects; Gene Expression Regulation - physiology; Human; Humans; Immunohistochemistry - methods; Intercellular Signaling Peptides and Proteins - pharmacology; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Nerve Tissue Proteins - metabolism; Neural progenitor; Neural stem cell; Neurons - drug effects; Neurosciences; Neurospheres; Neurovetenskaper; Parkinson's disease; Prosencephalon - cytology; Prosencephalon - embryology; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction - methods; Stem Cells - drug effects; Stem Cells - metabolism; Time Factors; Tyrosine 3-Monooxygenase - metabolism; Tyrosine hydroxylase; Ventral Tegmental Area - metabolism; Ventral Tegmental Area - transplantation
• ISSN: 0361-9230; 1873-2747; 1873-2747
• DOI: 10.1016/j.brainresbull.2006.07.001

Multipotent stem/progenitor cells derived from human first trimester forebrain can be expanded as free-floating aggregates, so called neurospheres. These cells can differentiate into neurons, astrocytes and oligodendrocytes. In vitro differentiation protocols normally yield γ-aminobutyric acid-immunoreactive neurons, whereas only few tyrosine hydroxylase (TH) expressing neurons are found. The present report describes conditions under which 4–10% of the cells in the culture become TH immunoreactive (ir) neurons within 24 h. Factors including acidic fibroblast growth factor (aFGF) in combination with agents that increase intracellular cyclic AMP and activate protein kinase C, in addition to a substrate that promotes neuronal differentiation appear critical for efficient TH induction. The cells remain THir after trypsinization and replating, even when their subsequent culturing takes place in the absence of inducing factors. Consistent with a dopaminergic phenotype, mRNAs encoding aromatic acid decarboxylase, but not dopamine-β-hydroxylase were detected by quantitative real time RT-PCR. Ten weeks after the cells had been grafted into the striatum of adult rats with unilateral nigrostriatal lesions, only very few of the surviving human neurons expressed TH. Our data suggest that a significant proportion of expandable human neural progenitors can differentiate into TH-expressing cells in vitro and that they could be useful for drug and gene discovery. Additional experiments, however, are required to improve the survival and phenotypic stability of these cells before they can be considered useful for cell replacement therapy in Parkinson's disease.