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

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

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SCHOLZ, Diana, Dominik PÖLTL, Andreas GENEWSKY, Matthias WENG, Tanja WALDMANN, Stefan SCHILDKNECHT, Marcel LEIST, 2011. Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line. In: Journal of Neurochemistry. 119(5), pp. 957-971. ISSN 0022-3042. eISSN 1471-4159. Available under: doi: 10.1111/j.1471-4159.2011.07255.x

@article{Scholz2011-12Rapid-18149, title={Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line}, year={2011}, doi={10.1111/j.1471-4159.2011.07255.x}, number={5}, volume={119}, issn={0022-3042}, journal={Journal of Neurochemistry}, pages={957--971}, author={Scholz, Diana and Pöltl, Dominik and Genewsky, Andreas and Weng, Matthias and Waldmann, Tanja and Schildknecht, Stefan and Leist, Marcel} }

Leist, Marcel Weng, Matthias Scholz, Diana Scholz, Diana Waldmann, Tanja Rapid, complete and large-scale generation of post-mitotic neurons from the human LUHMES cell line Weng, Matthias eng deposit-license Publ. in: Journal of Neurochemistry ; 119 (2011), 5. - S. 957-971 Genewsky, Andreas 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. 2012-02-01T08:19:06Z Waldmann, Tanja Leist, Marcel Genewsky, Andreas 2012-02-01T08:19:06Z Pöltl, Dominik Schildknecht, Stefan Pöltl, Dominik Schildknecht, Stefan 2011-12

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