title: Two methods of vitrification followed by in vitro culture of the ovine ovary: evaluation of the follicular development and ovarian extracellular matrix.
abstract: The aim of this study was to evaluate the influence of two vitrification techniques on the extra cellular matrix (ECM) and ovarian follicular development. The ovarian cortex was fragmented (9 mm(3)) and divided into six groups, viz. fresh control, cultured control, vitrified by the Ovarian Tissue Cryosystem (OTC) method, conventional solid surface vitrification (SSV) method, OTC/cultured and SSV/cultured. Follicles from all the fragments were analysed for morphology, development and viability. The ECM was evaluated based on the condition of collagen and reticular fibres and the immunolocalization of type I collagen and fibronectin. After 7 days of culture, the tissue vitrified by OTC revealed a higher percentage (p < 0.05) of morphologically normal (30.66%) and viable (60.00%) follicles when compared with those vitrified using the SSV technique (21.33% and 23.00%). In all the fragments cultured, regardless of the vitrification method, a significantly higher percentage of developing follicles was observed when compared with the non-cultured tissue. Analysis of the type I collagen showed increased immunostaining after the in vitro culture in the vitrified fragments. In conclusion, the OTC is better for preserving the follicular viability and morphology and maintaining the integrity of the extracellular matrix components of the ovine ovary.
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: F T
authors.initials: FT
authors.lastname: Bandeira
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: A A
authors.initials: AA
authors.lastname: Carvalho
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: S V
authors.initials: SV
authors.lastname: Castro
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: L F
authors.initials: LF
authors.lastname: Lima
authors.affiliation: Laboratory of Histology of Effects Caused by Poisons of Snakes and Plants, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: D A
authors.initials: DA
authors.lastname: Viana
authors.affiliation: Laboratory Veterinary Pathology, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: Jsam
authors.initials: J
authors.lastname: Evangelista
authors.affiliation: Laboratory of Neurophysiology, Institute of Biology Roberto Alcântara Gomes, State University of Rio de Janeiro, Rio de Janeiro-RJ, Brazil.
authors.forename: Mjs
authors.initials: M
authors.lastname: Pereira
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: C C
authors.initials: CC
authors.lastname: Campello
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: J R
authors.initials: JR
authors.lastname: Figueiredo
authors.affiliation: Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza-CE, Brazil.
authors.forename: Apr
authors.initials: A
authors.lastname: Rodrigues
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000502:physiology; D015925:Cryopreservation / Q000662:veterinary; D005109:Extracellular Matrix; D005260:Female; D006041:Goats; D007150:Immunohistochemistry / Q000662:veterinary; D006080:Ovarian Follicle / Q000166:cytology / Q000502:physiology*; D046509:Tissue Culture Techniques; D058989:Vitrification