title: In vitro culture of mouse preantral follicles using membrane inserts and developmental competence of in vitro ovulated oocytes.
abstract: To develop a reliable follicle culture system, mouse preantral follicles 150-200 microm in diameter were cultured individually for 5 or 6 days in membrane inserts or in droplets, and then induced to ovulate with hCG (Experiment 1). The nuclear maturation and developmental competence of the oocytes that ovulated from the follicles cultured in inserts were determined (Experiment 2). There was no significant difference between the two culture systems in the survival rate (83 and 77%). However, follicles cultured in inserts showed a higher ovulation rate (63%) than those cultured in droplets (39%, P<0.05). About 80% of the oocytes that ovulated from the follicles cultured in inserts were at the metaphase II stage. After in vitro fertilization, 75 and 48% of in vitro ovulated oocytes cleaved and developed into blastocysts, respectively. These results demonstrate that the insert culture system is superior to the droplet culture system in terms of follicular growth and ovulation, and can be used to investigate the growth and ovulation of follicles in vitro.
authors.affiliation: Laboratory of Theriogenology, Department of Veterinary Clinical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
authors.forename: Ahmed Abdel Gadir
authors.initials: AA
authors.lastname: Adam
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Seiji
authors.initials: S
authors.lastname: Katagiri
authors.forename: Masashi
authors.initials: M
authors.lastname: Nagano
mesh_terms: D000818:Animals; D002470:Cell Survival; D005260:Female; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000166:cytology* / Q000502:physiology*; D009924:Organ Culture Techniques / Q000379:methods*; D006080:Ovarian Follicle / Q000166:cytology* / Q000254:growth & development / Q000502:physiology*; D010060:Ovulation