Enhanced differentiation capacity of parthenogenetic embryonic stem cells via incorporation of non-growing oocyte genomes in mouse.

Objectives: To evaluate the in vivo developmental and therapeutic potential of a novel parthenogenetic embryonic stem cell line (NF-pES), which contains genomes from both non-growing and grown oocytes.

Methods: NF-pES cells were injected into mouse blastocysts to generate chimeric mice, and their contribution to various tissues was assessed. Skeletal muscle differentiation potential was examined through teratoma assays and analysis of muscle tissue in chimeric mice. For therapeutic assessment, a skeletal muscle injury model was established by cardiotoxin and irradiation treatment of the tibialis anterior muscle. NF-pES-derived precursor cells, obtained through in vitro induction and differentiation, were transplanted into the injured muscle.

Results: Notably, NF-pES cells contributed extensively to multiple somatic lineages in chimeric mice, with high levels of chimerism observed in the heart (83.36%) and bone marrow (50.44%). These levels are comparable to those achieved with embryonic stem cells derived from fertilized embryos. Importantly, NF-pES cells demonstrated robust myogenic differentiation capacity, as evidenced by their contribution to skeletal muscle tissues in both teratoma formation assays and in vivo chimeric muscle integration. Following in vitro induction, NF-pES-derived precursors were transplanted into the injured tibialis anterior muscle of recipient mice to assess their regenerative potential in vivo. One month after transplantation, immunohistochemical analysis confirmed the successful engraftment of donor-derived cells within the host muscle tissue. These donor-derived cells expressed markers of terminal myogenic differentiation and were incorporated into mature skeletal muscle fibers.

Conclusions: NF-pES cells exhibit strong developmental capacity and therapeutic potential for skeletal muscle regeneration, suggesting their value in future regenerative medicine applications.