Parthenogenetic embryonic stem cells derived from cryopreserved newborn mouse ovaries: a new approach to autologous stem cell therapy

ObjectiveTo evaluate whether cryopreserved newborn mouse ovaries can generate sufficient numbers of parthenogenetic mouse embryonic stem (pmES) cells for autologous stem cell therapy.DesignProspective study.SettingReproductive clinic of Xinhua Hospital in Shanghai.Animal(s)Kunming, C57BL/6J, BALB/c, and NOD-SCID mice.Intervention(s)Cryopreserved newborn mouse ovaries were thawed, grafted into immunodeficient mice, treated with pregnant mare serum gonadotropin to promote follicular maturation, and collected oocytes activated in vitro to generate parthenogenetic embryonic stem cells.Main Outcome Measure(s)Preimplantation development and stem cell characterization.Result(s)This new protocol yielded a large number of oocytes from cryopreserved ovaries over a long period. These oocytes were used to derive pmES cell lines, which expressed embryonic stem cell–specific markers and differentiated into embryoid bodies in vitro and teratomas in vivo. The pmES cell line was propagated in an undifferentiated state for more than 30 passages and maintained a diploid karyotype.Conclusion(s)The pmES cells lines established by our protocol exhibited the same degree of pluripotency as standard embryonic stem cell lines. This approach may be used for exploring autologous stem cell therapies.