title: Comparison of three in vitro culture systems for maturation of early preantral mouse ovarian follicles.
abstract: The aim of this study was to compare three different culture systems for in vitro follicular growth and oocyte maturation in ovarian follicles of mice in order to assess the technique with the optimal growth and improved rate of meiotic maturation. The three systems tested were culture under oil, on a hydrophobic membrane and on agar respectively. Early preantral follicles were cultured for 12 days in alpha-MEM GlutaMAX medium. Follicular growth, oocyte meiotic maturation, oocyte extrusion, atresia and estradiol production were analysed. Follicular development showed two phases in the three systems, with slow growth before day 5 and subsequent acceleration. The percentage of follicles transferred into oocyte maturation medium was significantly higher after culture under oil. The proportion of oocytes that achieved nuclear maturation (metaphase II) was higher when follicles were cultured under oil or on a hydrophobic membrane than on agar. Our results support the use of culture under oil for in vitro follicular growth from the early preantral stage in order to obtain metaphase II oocytes. Fertilization ability of these oocytes and the capacity to obtain healthy mice in a reproducible manner warrants further investigation.
authors.affiliation: Cochin Hospital, University of Paris V, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Mousset-Simeón
authors.forename: Pierre
authors.initials: P
authors.lastname: Jouannet
authors.forename: Laëtitia
authors.initials: L
authors.lastname: Le Cointre
authors.forename: Christiane
authors.initials: C
authors.lastname: Coussieu
authors.forename: Catherine
authors.initials: C
authors.lastname: Poirot
mesh_terms: D000362:Agar; D000818:Animals; D003470:Culture Media; D005260:Female; D008567:Membranes, Artificial; D051379:Mice; D008810:Mice, Inbred C57BL; D008899:Mineral Oil; D009865:Oocytes / Q000166:cytology / Q000254:growth & development*; D009930:Organic Chemicals; D006080:Ovarian Follicle / Q000166:cytology / Q000254:growth & development*; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D003470:Culture Media; D008567:Membranes, Artificial; D009930:Organic Chemicals; C420642:alpha minimal essential medium; D008899:Mineral Oil; D000362:Agar