Androgenetic Embryonic Stem Cells form Neural Progenitor Cells in vivo And in vitro

¹ Institut für Medizinische Strahlenkunde und Zellforschung (MSZ), Universität Würzburg, Würzburg, Germany
² Center for Animal Transgenesis and Germ Cell Research, New Bolton Center, University of Pennsylvania, Kennett Square PA, USA

^* To whom correspondence should be addressed. E-mail: albrecht.mueller{at}mail.uni-wuerzburg.de.

	Abstract

Uniparental zygotes with two paternal (androgenetic: AG) or two maternal (gynogenetic: GG; parthenogenetic: PG) genomes are not able to develop into viable offspring but can form blastocysts from which embryonic stem cells (ESCs) can be derived. Although some aspects of the in vitro and in vivo differentiation potential of PG and GG ESCs of several species have been studied, the developmental capacity of AG ESCs is much less clear. Here, we investigate the potential of murine AG ESCs to undergo neural differentiation. We observed that AG ESCs differentiate in vitro into pan-neural progenitor cells (pnPCs) that further give rise to cells that express neuronal- and astroglial-specific markers. Neural progeny of in vitro differentiated AG ESCs exhibited fidelity of expression of six imprinted genes analyzed with the exception of Ube3a. Bisulfite-sequencing for two imprinting control regions suggested that pnPCs predominantly maintained their methylation pattern. Following blastocyst injection of AG and biparental (normal fertilized: N) ESCs we found widespread and evenly distributed contribution of ESC-derived cells in both AG and N chimeric early fetal brains. AG and N ESC-derived cells isolated from chimeric fetal brains by fluorescence activated cell sorting (FACS) exhibited similar neurosphere initiating cell frequencies and neural multi-lineage differentiation potential. Our results indicate that AG ESC-derived neural progenitor/stem cells do not differ from N neural progenitor/stem cells in their self-renewal and neural multi-lineage differentiation potential.