title: Extended ex vivo culture of fresh and cryopreserved whole sheep ovaries.
abstract: We describe an original perfusion system for the culture of whole ovine ovaries for up to 4 days. A total of 33 ovaries were divided into six groups: control (n=6), not perfused and fixed; Groups SM72 and SM72-FSH (n=6 each), perfused with a simple medium for 72h with or without FSH; Groups CM96 and CM96-FSH (n=6 each), perfused with a complex medium for 96h with or without FSH; Group CM96-FSH-cryo, (n=3) cryopreserved and perfused for 96h with Group CM96-FSH medium. Depending on the medium used, morphological parameters of cultured ovaries differed from fresh organs after 72 (SM72, SM72-FSH) or 96 (CM96, CM96-FSH) h of perfusion. Oestradiol and progesterone were secreted in all groups but FSH had an effect only on Group CM96-FSH, stimulating continued oestradiol secretion 10 times higher than in all other groups. Morphological parameters and hormone secretion of cryopreserved ovaries were not different from fresh controls. This method enables the culture of whole ovaries for up to 4 days, the time required in vivo for 0.5-mm follicles to grow to 2.2mm and then for these follicles to reach the ovulatory size of 4mm or more. It could be used as a research tool or to complement current techniques for preserving female fertility.
authors.affiliation: Institute for Genetic and Biomedical Research, National Research Council, Milan, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Maffei
authors.affiliation: Department of Veterinary Medical Sciences, Università degli Studi di Bologna, Via Tolara di Sopra, 50 40064 Ozzano dell'Emilia, Italy.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Galeati
authors.affiliation: Laboratory of Biomedical Embryology, Università degli Studi di Milano, Italy.
authors.forename: Georgia
authors.initials: G
authors.lastname: Pennarossa
authors.affiliation: Laboratory of Biomedical Embryology, Università degli Studi di Milano, Italy.
authors.forename: Tiziana A L
authors.initials: TAL
authors.lastname: Brevini
authors.affiliation: Present address: Department of Health, Animal Science and Food Safety, Università degli Studi di Milano via Celoria 10, 20133 Milan, Italy.
authors.forename: Fulvio
authors.initials: F
authors.lastname: Gandolfi
mesh_terms: D000818:Animals; D015925:Cryopreservation; D004958:Estradiol / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone; D009924:Organ Culture Techniques; D010053:Ovary / Q000502:physiology*; D011374:Progesterone / Q000378:metabolism; D012756:Sheep
chemical_list: D011374:Progesterone; D004958:Estradiol; D005640:Follicle Stimulating Hormone