Two Methods of Vitrification Followed by In Vitro Culture of the Ovine Ovary: Evaluation of the Follicular Development and Ovarian Extracellular Matrix

• Published in: Reproduction in domestic animals Vol. 50; no. 2; pp. 177 - 185
• Main Authors: Bandeira, FT, Carvalho, AA, Castro, SV, Lima, LF, Viana, DA, Evangelista, JSAM, Pereira, MJS, Campello, CC, Figueiredo, JR, Rodrigues, APR
• Format: Journal Article
• Language: English
• Published: Germany P. Parey Scientific Publishers 01-04-2015; Blackwell Publishing Ltd
• Subjects: animal ovaries; Animal reproduction; Animals; Cattle; Cell culture; Cell Proliferation - physiology; Collagen; Collagen (type I); cortex; Cryopreservation - veterinary; Extracellular Matrix; Female; Fibronectin; fibronectins; Follicles; follicular development; Fragmentation; Fragments; Goats; Immunohistochemistry - veterinary; in vitro culture; Morphology; Ovarian Follicle - cytology; Ovarian Follicle - physiology; sheep; Tissue analysis; Tissue culture; Tissue Culture Techniques; Viability; Vitrification
• ISSN: 0936-6768; 1439-0531
• DOI: 10.1111/rda.12463

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.