Comparative characterization of four mouse parthenogenetic embryonic stem (pES) cell lines

Abstract

Derivation of embryonic stem (ES) cells from parthenogenetic embryos represents a possible alternative approach to create histocompatible cells for regenerative medicine. The objectives of this study were to establish mouse parthenogenetic ES (pES) cell lines from parthenogenetically-derived blastocysts as a model system for human and animal research and to examine pluripotency differences among the pES cell lines. We are able to report the successful establishment of four pluripotent pES cell lines from blastocysts of parthenogenetic origin (22% efficiency of pES cell line establishment). Four pES cell lines (pES#1-4) exhibited a typical ES cell morphology and expression of key pluripotency markers (ALP, Oct4, Nanog and SSEA-1). Three of the four pES cell lines have shown a high percentage of normal karyotype during long-term culture. Variability in the in vitro differentiation potential into cell types of the 3 germ layers was observed among the different pES cell lines. Three of these (pES#1-3) exhibited a higher efficiency towards endo-mesoderm differentiation, strongly expressed differentiation markers towards endomesoderm lineage (α-fetoprotein; Flk-1; PECAM and collagen IV) than pES#4. Differentiation towards cardiac cells resulted in all cell lines 33-100% of spontaneous beating cell clusters/well. Furthermore, following injection into blastocysts pES#1 cells differentiated successfully in vivo into chimeric mice with an efficiency of 75% (three chimeras of four newborns). In conclusion, our results have demonstrated that there are major differences among pES lines in their differentiation ability in vitro and that it was possible to generate chimera forming pES cell lines in mouse.