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Comparison of three in vitro culture systems for maturation of early preantral mouse ovarian follicles

Published online by Cambridge University Press:  20 July 2005

Nathalie Mousset-Siméon ,
Pierre Jouannet ,
Laëtitia Le Cointre ,
Christiane Coussieu  and
Catherine Poirot
Nathalie Mousset-Siméon
Affiliation:
GREFH, Reproduction Biology Laboratory, Cochin Hospital – University of Paris V, 123 boulevard de Port-Royal, 75679 Paris Cedex 14, France. Reproductive Biology Laboratory, Rouen University – Hospital Charles Nicolle, 1 rue de Germont, 76031 Rouen Cedex, France.
Pierre Jouannet
Affiliation:
GREFH, Reproduction Biology Laboratory, Cochin Hospital – University of Paris V, 123 boulevard de Port-Royal, 75679 Paris Cedex 14, France.
Laëtitia Le Cointre
Affiliation:
Biochemistry Laboratory, Hôtel Dieu Hospital of Paris, 1 place du parvis Notre-Dame, 75181 Paris Cedex 4, France.
Christiane Coussieu
Affiliation:
Biochemistry Laboratory, Hôtel Dieu Hospital of Paris, 1 place du parvis Notre-Dame, 75181 Paris Cedex 4, France.
Catherine Poirot
Affiliation:
GREFH, Reproduction Biology Laboratory, Cochin Hospital – University of Paris V, 123 boulevard de Port-Royal, 75679 Paris Cedex 14, France. Reproductive Biology Laboratory, Pitié-Salpétrière Hospital of Paris, 83 boulevard de l'hôpital, 75651 Paris Cedex 13, France.
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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 α-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.

Keywords

CultureEstradiolMaturationMouseOocytePreantral follicle
Type
Research Article
Information
Zygote , Volume 13 , Issue 2 , May 2005 , pp. 167 - 175
Copyright
2005 Cambridge University Press

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