Parthenogenetic embryonic stem cells derived from cryopreserved newborn mouse ovaries: a new approach to autologous stem cell therapy.
OBJECTIVE
To evaluate whether cryopreserved newborn mouse ovaries can generate sufficient numbers of parthenogenetic mouse embryonic stem (pmES) cells for autologous stem cell therapy.


METHODS
Prospective study.


METHODS
Reproductive clinic of Xinhua Hospital in Shanghai.


METHODS
Kunming, C57BL/6J, BALB/c, and NOD-SCID mice.


METHODS
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.


METHODS
Preimplantation development and stem cell characterization.


RESULTS
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.


CONCLUSIONS
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.
Xing
Fengying
F
Center for Developmental Biology, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Fang
Zhenfu
Z
Qin
Bolin
B
Li
Yao
Y
Hou
Jian
J
Chen
Xuejin
X
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods; D053595:Embryonic Stem Cells / Q000502:physiology*; D005260:Female; D007621:Karyotyping; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009865:Oocytes / Q000502:physiology; D009866:Oogenesis / Q000502:physiology; D010053:Ovary / Q000637:transplantation; D010312:Parthenogenesis / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods*; D014182:Transplantation, Autologous