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Vitrification of ovarian tissue of Brazilian North‐eastern donkeys ( Equus asinus ) using different cryoprotectants
Author(s) -
Lopes Kátia Regina F.,
Praxedes Erica Camila G.,
Campos Livia B.,
Bezerra Marcelo B.,
Lima Gabriela L.,
Saraiva Márcia Viviane A.,
Silva Alexandre R.
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.13203
Subject(s) - cryoprotectant , vitrification , dimethyl sulfoxide , andrology , follicular phase , ethylene glycol , cryopreservation , biology , chemistry , zoology , endocrinology , embryo , medicine , organic chemistry , microbiology and biotechnology
Contents The aim of this study was to assess a vitrification protocol for asinine ovarian tissue, to preserve preantral follicles using different cryoprotectant solutions, composed of various concentrations ( EG 3 M or 6 M) of dimethyl sulfoxide or ethylene glycol isolate, or as a combination ( DMSO 3 M + EG 3 M). Ten pairs of ovaries from Brazilian north‐eastern breed jennies were obtained through videolaparoscopy, and cortical fragments were submitted to a solid‐surface vitrification ( SSV ) using each cryoprotectant solution. The ovarian tissue was evaluated for follicular morphology and viability, DNA integrity ( TUNEL technique) and the presence of nucleolar organizing regions in granulosa cells (Ag NOR technique). After thawing, the percentage of normal preantral follicles was significantly reduced in the vitrified ovarian tissue fragments compared to the fresh control ( p < 0.05). When comparing treatments, the use of DMSO 3 M (81.7 ± 37.5%), EG 3 M (83.7 ± 27.4%) and the combination of both DMSO 3 M + EG 3 M (81.8 ± 46.8%) allowed a greater percentage of follicular survival in contrast to DMSO 6 M (69.8 ± 16.5%) and EG 6 M (72.3 ± 18.0%; p < 0.05). When vitrified using the DMSO + EG combination, a higher percentage (62.5 ± 29.1%) of viable follicles (trypan blue) was observed in relation to the other vitrification treatments ( p < 0.05). The TUNEL technique identified that all treatments tested showed DNA fragmentation in the follicular cells, except in the case of the DMSO 3 M + EG 3 M treatment. When evaluating the presence of NOR s, no significant differences were observed in the amount of NOR s between the fresh and vitrified groups using DMSO 3 M + EG 3 M ( p > 0.05). We concluded that the combination DMSO 3 M + EG was more efficient for the vitrification of ovarian tissue taken from Equus asinus , allowing adequate preservation of PAF s morphology, viability, DNA integrity and cell proliferative capacity.