TY - JOUR A1 - Ahmad, Ruhel A1 - Wolber, Wanja A1 - Eckardt, Sigrid A1 - Koch, Philipp A1 - Schmitt, Jessica A1 - Semechkin, Ruslan A1 - Geis, Christian A1 - Heckmann, Manfred A1 - Brüstle, Oliver A1 - McLaughlin, John K. A1 - Sirén, Anna-Leena A1 - Müller, Albrecht M. T1 - Functional Neuronal Cells Generated by Human Parthenogenetic Stem Cells JF - PLoS One N2 - Parent of origin imprints on the genome have been implicated in the regulation of neural cell type differentiation. The ability of human parthenogenetic (PG) embryonic stem cells (hpESCs) to undergo neural lineage and cell type-specific differentiation is undefined. We determined the potential of hpESCs to differentiate into various neural subtypes. Concurrently, we examined DNA methylation and expression status of imprinted genes. Under culture conditions promoting neural differentiation, hpESC-derived neural stem cells (hpNSCs) gave rise to glia and neuron-like cells that expressed subtype-specific markers and generated action potentials. Analysis of imprinting in hpESCs and in hpNSCs revealed that maternal-specific gene expression patterns and imprinting marks were generally maintained in PG cells upon differentiation. Our results demonstrate that despite the lack of a paternal genome, hpESCs generate proliferating NSCs that are capable of differentiation into physiologically functional neuron-like cells and maintain allele-specific expression of imprinted genes. Thus, hpESCs can serve as a model to study the role of maternal and paternal genomes in neural development and to better understand imprinting-associated brain diseases. KW - methylation KW - derivation KW - blastocysts KW - pluripotent KW - differentiation KW - lines KW - brain development KW - in-vitro KW - mice KW - specification Y1 - 2012 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-130268 VL - 7 IS - 8 ER - TY - THES A1 - Wolber, Wanja Andrej T1 - Neuronales Differenzierungspotential muriner androgenetischer Embryonaler- Stammzellen in „vitro“ und in „vivo“ T1 - Neural Differentiation of Androgenetic Murine ES Cell-Derived Neural Progenitor Cells in vitro and in vivo N2 - In dieser Arbeit wurde gezeigt, dass aus uniparentalen, embryonalen Stammzellen mit fehlender maternal geprägter Genexpression (AG-Zellen) differenzierte neuronale Progenitorzellen (pNPCs) eine ähnliche neuronale Kapazität wie wildtypische Progenitorzellen haben. Sie bilden nach histomorphologischen Kriterien in vitro adulte Neurone mit Ausbildung eines synaptischen Netzwerks. In elektrophysiologischen PatchClamp- Untersuchungen wurde gezeigt, dass diese Zellen, ähnlich dem wildtypischen Pendant, spannungsabhängige Natrium- und Kaliumkanälen besitzen, ein negatives Membranpotential haben und bei Stimulation mit repetitiven Aktionspotentialen reagieren. Nach Transplantation in einem Schädel-Hirn- Trauma-Modell konnten nach drei Monaten in vivo Donorzellen mit neuraler Morphologie und der Expression von jungen, neuronalen und glialen Proteinen gefunden werden. Die Teratombildung ist im Vergleich zum Wildtyp unverändert, eine maligne Entartung mit invasivem Wachstum oder ausgedehnter Metastasierung konnte nicht gefunden werden. Aus AG-Zellen generierte neuronale Progenitorzellen sind ein starkes Instrument, um neuronale genomische Prägung zu untersuchen. Außerdem könnte die regenerative Kapazität für eine patientenspezifische Zellersatztherapie genutzt werden. N2 - We have shown that uniparental maternal [AG-origin] embryonic stem cells can give rise to neural progenitor cells [pNPCs] and have a similar ability to form neural tissue compared to wildtype progenitor cells. In vitro they can develop into adult neurons that form a synaptic network. In patch clamp experiments it could be shown that these cells produce a similar amount of voltage gated sodium- and potassium channels, have a negative membrane potential and form multiple action potentials after depolarisation. Three month after transplantation donor cells with expression of early neural development could be found in a transplantation modell afterbrain trauma. The frequency of teratoma forming does not vary compared to wildtype cells. Neural progenitor cells derived from AG-ES cells are a strong tool to study neural genomic imprinting. Their regenerative capacity could be used for a patient specific cell replacement therapy. KW - Uniparentale Stammzellen Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-165929 ER -