Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line

Authors

  • Diana Scholz,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
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    • These authors contributed equally to this study.

  • Dominik Pöltl,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
    2. Konstanz Research School Chemical Biology, University of Konstanz, Konstanz, Germany
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    • These authors contributed equally to this study.

  • Andreas Genewsky,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
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  • Matthias Weng,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
    2. Konstanz Research School Chemical Biology, University of Konstanz, Konstanz, Germany
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  • Tanja Waldmann,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
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  • Stefan Schildknecht,

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
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  • Marcel Leist

    1. Doerenkamp-Zbinden Chair for in vitro Toxicology and Biomedicine, University of Konstanz, Konstanz, Germany
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Address correspondence and reprint requests to Diana Scholz and Dominik Pöltl, University of Konstanz, Universitätsstraße 10, Postbox M657, D-78457 Konstanz, Germany. E-mail: diana.scholz@uni-konstanz.de; dominik.poeltl@uni-konstanz.de

Abstract

J. Neurochem. (2011) 119, 957–971.

Abstract

We characterized phenotype and function of a fetal human mesencephalic cell line (LUHMES, Lund human mesencephalic) as neuronal model system. Neurodevelopmental profiling of the proliferation stage (d0, day 0) of these conditionally-immortalized cells revealed neuronal features, expressed simultaneously with some early neuroblast and stem cell markers. An optimized 2-step differentiation procedure, triggered by shut-down of the myc transgene, resulted in uniformly post-mitotic neurons within 5 days (d5). This was associated with down-regulation of some precursor markers and further up-regulation of neuronal genes. Neurite network formation involved the outgrowth of 1–2, often > 500 μm long projections. They showed dynamic growth cone behavior, as evidenced by time-lapse imaging of stably GFP-over-expressing cells. Voltage-dependent sodium channels and spontaneous electrical activity of LUHMES continuously increased from d0 to d11, while levels of synaptic markers reached their maximum on d5. The developmental expression patterns of most genes and of the dopamine uptake- and release-machinery appeared to be intrinsically predetermined, as the differentiation proceeded similarly when external factors such as dibutyryl-cAMP and glial cell derived neurotrophic factor were omitted. Only tyrosine hydroxylase required the continuous presence of cAMP. In conclusion, LUHMES are a robust neuronal model with adaptable phenotype and high value for neurodevelopmental studies, disease modeling and neuropharmacology.

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