Overexpression of insulin-like growth factor-II in mouse embryonic stem cells promotes myogenic differentiation.

Embryonic stem (ES) cells derived from androgenetic or parthenogenetic mouse embryos are important tools for studying the roles of imprinted genes in early development. Androgenetic ES cells have been shown to preferentially differentiate into the myogenic lineage both in vitro and after formation of teratocarcinomas in vivo. To clarify if the maternally imprinted Igf2 gene which is expected to be overexpressed in androgenetic ES cells is sufficient to induce myogenic differentiation, R1 ES cells were transfected with human IGF-II expression vectors. Stable ES cell clones exhibiting human IGF-II mRNA and protein expression were studied vs ES cell clones without IGF-II overexpression in a standard in vitro differentiation system involving culture in "hanging drops" and observation of differentiation of the recovered embryoid bodies (EBs). EBs derived from IGF-II overexpressing ES cells showed stimulated myogenic differentiation evident by the appearance of myoblasts already 3 days after plating and by higher levels of skeletal muscle-specific transcripts (myf5, myoD, myogenin) at earlier stages. Our study demonstrates for the first time that overexpression of IGF-II enhances and accelerates myogenic differentiation of ES cells, which has implications for ES cell-derived tissue engineering.