Derivation of developmentally competent oocytes by the culture of preantral follicles retrieved from adult ovaries: maturation, blastocyst formation, and embryonic stem cell transformation.
OBJECTIVE
To determine whether the preantral follicles in adult ovaries can generate developmentally competent oocytes after in vitro culture.


METHODS
Prospective, animal-model study.


METHODS
Gamete and Stem Cell Biotechnology Laboratory, Seoul National University, Seoul, Korea.


METHODS
B6CBAF1 mice.


METHODS
Preantral follicles collected from 8-week-old mice were cultured in vitro.


METHODS
Follicle development, embryogenesis, and embryonic stem cell characterization.


RESULTS
A mean of 50.3 preantral follicles were retrieved from one adult animal, which is significantly less than the number (88.7 follicles) retrieved from a prepubertal female. Extension of the culture period greatly improved oocyte maturation; increased follicular growth to the pseudo-antral (89%-91% vs. 32%) or mature oocyte stage (65%-77% vs. 13%) was observed after 12 or 13 days of culture compared with 9 days of culture. Blastocyst formation after parthenogenesis was detected in only one case; in comparison, the use of IVF yielded a large number of embryos that developed into blastocysts. A mean of 14.7 intrafollicular oocytes per animal were produced after 13 days of culture, and 41% of those developed into blastocysts after IVF. Embryonic stem cell-like colonies were established by subculturing the inner cell mass cells from the blastocysts.


CONCLUSIONS
Developmentally competent oocytes can be generated by culturing adult preantral follicles. These results may help increase the feasibility of follicle culture systems.
Kim
In Wook
IW
Gamete and Stem Cell Biotechnology Laboratory, Department of Agricultural Biotechnology, Seoul National University, Seoul, South Korea.
Gong
Seung Pyo
SP
Yoo
Cho Rong
CR
Choi
Jun Hee
JH
Kim
Dae Yong
DY
Lim
Jeong Mook
JM
D000367:Age Factors; D000818:Animals; D001755:Blastocyst / Q000502:physiology; D018929:Cell Culture Techniques; D002461:Cell Line, Transformed; D002469:Cell Separation / Q000379:methods; D002471:Cell Transformation, Neoplastic / Q000473:pathology; D002478:Cells, Cultured; D047108:Embryonic Development / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000473:pathology* / Q000502:physiology; D005240:Feasibility Studies; D005260:Female; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D006080:Ovarian Follicle / Q000166:cytology*; D010053:Ovary / Q000166:cytology; D010312:Parthenogenesis / Q000502:physiology*