Establishment of autologous embryonic stem cells derived from preantral follicle culture and oocyte parthenogenesis.

OBJECTIVE: To evaluate whether autologous embryonic stem cells can be established without generating clone embryos.
DESIGN: Prospective model study.
SETTING: Gamete and stem cell biotechnology laboratory in Seoul National University, Seoul, Korea. ANIMAL(S): F1 hybrid B6D2F1 mice. INTERVENTION(S): Preantral follicles were cultured, and oocytes matured in the follicles were parthenogenetically activated. MAIN OUTCOME MEASURE(S): Preimplantation development and stem cell characterization. RESULT(S): More intrafollicular oocytes that were retrieved from secondary follicles matured and developed into blastocysts after parthenogenesis than those that were retrieved from primary follicles. Of those 35 blastocysts derived from 193 parthenotes, one line of colony-forming cells was established from the culturing of early secondary follicles. The established cells were positive for embryonic stem cell-specific markers and had normal diploid karyotype and telomerase activity. They differentiated into embryoid bodies in vitro and teratomas in vivo. Inducible differentiation of the established cells into neuronal lineage cells also was possible. CONCLUSION(S): Autologous embryonic stem cells can be established by preantral follicle culture and oocyte parthenogenesis. A combined technique of follicle culture and oocyte parthenogenesis that does not use developmentally competent oocytes has the potential to replace somatic cell nuclear transfer for autologous cell therapy.