The Thai Journal of Veterinary Medicine


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

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