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Vitrification of cat ovarian tissue: Does fragment size matters?
Author(s) -
Gorricho Camila M.,
Tavares Mariana R.,
Apparício Maricy,
FonsecaAlves Carlos E.,
Macente Beatrice I.,
Mansano Cleber F.M.,
Toniollo Gilson H.
Publication year - 2018
Publication title -
reproduction in domestic animals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.546
H-Index - 66
eISSN - 1439-0531
pISSN - 0936-6768
DOI - 10.1111/rda.13357
Subject(s) - andrology , cryoprotectant , vitrification , immunohistochemistry , apoptosis , ovarian tissue , biology , cryopreservation , caspase 3 , ovary , embryo , programmed cell death , endocrinology , immunology , medicine , microbiology and biotechnology , biochemistry
Contents The aim of the present study was to investigate whether or not the size of the ovarian fragment influences its resistance to cryostorage. For that purpose, ovaries were collected from 34 queens (various breeds, age 1–5 year) by routine ovariectomy, transported to the laboratory and then sectioned in different sizes (3 mm × 3 mm × 3 mm, 5 mm × 3 mm × 3 mm and 7 mm × 3 mm × 3 mm) and randomly assigned to a control ( GC 3, GC 5 and GC 7, respectively) or vitrified ( GV 3, GV 5 and GV 7, respectively) groups. Vitrified‐warmed fragments were evaluated by histomorphology and immunohistochemistry (for apoptotic rates by using cleaved caspase‐3). Histological examination reveals that 72.97% of the follicles in GV 3 and 72.58% in GV 5 were normal while only 42.86% of the follicles in GV 7. The main morphological alteration presented in all groups was a detachment of the epithelial cells. Similarly, immunohistochemistry evaluation using caspase 3 revealed a small proportion of apoptotic cells in GV 3 (8.43%) while in GV 7 30.43% of the cells expressed cleaved caspase‐3. These findings indicate that fragments sectioned in 3 mm × 3 mm × 3 mm (27 mm 3 ) seem more adequate for perfusion of the cryoprotectant, causing less damage to the cell after vitrification‐warming.