Two Methods of Vitrification Followed by In Vitro Culture of the Ovine Ovary: Evaluation of the Follicular Development and Ovarian Extracellular Matrix |
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| Authors: | FT Bandeira AA Carvalho SV Castro LF Lima DA Viana JSAM Evangelista MJS Pereira CC Campello JR Figueiredo APR Rodrigues |
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| Institution: | 1. Laboratory of Manipulation of Oocytes and Preantral Follicles, Veterinary of Faculty, State University of Ceará, Fortaleza‐CE, Brazil;2. Laboratory of Histology of Effects Caused by Poisons of Snakes and Plants, Veterinary of Faculty, State University of Ceará, Fortaleza‐CE, Brazil;3. Laboratory Veterinary Pathology, Veterinary of Faculty, State University of Ceará, Fortaleza‐CE, Brazil;4. Laboratory of Neurophysiology, Institute of Biology Roberto Alcantara Gomes, State University of Rio de Janeiro, Rio de Janeiro‐RJ, Brazil |
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| 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 mm3) 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. |
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