@phdthesis{Dinger2008,
  author    = {Dinger, Timo Christoph},
  title     = {Neural Differentiation Potential of Murine Androgenetic Embryonic Stem Cells},
  url       = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-36215},
  school      = {Universit{\"a}t W{\"u}rzburg},
  year      = {2008},
  abstract  = {Uniparental zygotes with two genomes from the same sex can be established from fertilised oocytes after pronuclear exchange. They contain two maternal (gynogenetic; GG) or paternal (androgenetic; AG) pronuclei and are not competent to develop into viable offspring but they can form blastocysts from which embryonic stem cells (ES cells) can be derived. The developmental potential of uniparental ES cells is not fully investigated. The restricted developmental potential of uniparental cells is cell-intrinsic and probably reflects the different roles maternal and paternal genomes play during development. Following blastocyst injection, both GG and AG ES cells show biased and parent-of-origin-specific chimaera formation. While the in vitro and in vivo neural differentiation potential of GG ES cells is well characterised the neural developmental potential of AG ES cells is less clear. In an earlier study the group of K. John McLaughlin reported that AG and GG ES cell-derived hematopoietic stem cells conveyed long-term, multi-lineage hematopoietic engraftment with no associated pathologies (Eckardt et al., 2007). The aim of this study was to investigate the potential of AG uniparental murine ES cells to differentiate in vitro and in vivo into neural progenitor / stem cells and further into neurons, astro- and oligodendroglia in comparison to GG and biparental (normal fertilised; N) ES cells. Uniparental and biparental ES cells were obtained from K. John McLaughlin's group and a cell culture system was established to expand uniparental (AG, GG) and biparental N ES cells on murine embryonic fibroblasts (MEF). A multistep-protocol was used to differentiate ES cells towards pan-neural progenitor cells and neuronal and glial cell types (Br{\"u}stle et al., 1997). The ability of terminal neural differentiation in vitro was analysed by fluorescence microscopy using neuronal and glial lineage markers. In parallel, eGFP+ AG or N ES cells were injected into blastocysts prior to their transfer into foster mothers. At E12.5 and E14.5, embryos were isolated, forebrains were dissected and by means of fluorescence activated cell sorting (FACS) eGFP+ donor cells were isolated from chimeric brains. Both eGFP+ donor and corresponding eGFP- blastocyst-derived brain cells were expanded and analyses of differentiation potential and self-renewal capacity were performed. Also, cryosections of E12.5 chimeric brains were analysed for donor contribution to the neuronal lineage by immunofluorescence microscopy. Here it is described that following in vitro differentiation, AG pan-neural progenitor cells have similar abilities to differentiate into neuronal and glial lineages as GG and N pan-neural progenitor cells. In cryosections of E12.5 chimeric brains no differences in brain engraftment and formation of immature neuronal cells between uniparental AG and N donor cells were detected. AG and N ES cell-derived cells isolated from chimeric foetal brains by FACS exhibited similar neurosphere initiating cell frequencies and neural multi-lineage differentiation potential. Therefore, the data of this study suggest that the previously described differences in the in vivo engraftment pattern of uniparental inner cell mass (ICM) cells in foetal brains (Keverne et al., 1996) are not primarily due to limitations in the proliferation or differentiation properties of uniparental neural progenitor cells. The results presented here indicate that AG ES cell-derived neural progenitor / stem cells did not differ from N neural progenitor / stem cells in their self-renewal and their neural multi-lineage differentiation potential. Also AG ES cell-derived cells contributed to developing brains at early foetal developmental stages showing a widespread and balanced distribution in chimeric brains. AG brain cells form neurospheres with self-renewal and neural differentiation capacity similar to N ES cell-derived brain cells. Thus, the data of this study together indicate that the neural developmental potential in vivo and in vitro of AG and N ES cells does not differ.},
  subject      = {Stammzelle},
  language  = {en}
}