TY - JOUR
T1 - Use of synthetic polymers improves the quality of vitrified caprine preantral follicles in the ovarian tissue
AU - Montano Vizcarra, Diego Alberto
AU - Pinto Silva, Yago
AU - Bezerra Bruno, Jamily
AU - Calado Brito, Danielle Cristina
AU - Dipaz Berrocal, Deysi
AU - Mascena Silva, Luciana
AU - Gaudencio dos Santos Morais, Maria Luana
AU - Alves, Benner Geraldo
AU - Alves, Kele Amaral
AU - Weber Santos Cibin, Francielli
AU - Figueiredo, José Ricardo
AU - Zelinski, Mary B.
AU - Ribeiro Rodrigues, Ana Paula
N1 - Publisher Copyright:
© 2019 Elsevier GmbH
PY - 2020/2
Y1 - 2020/2
N2 - The aim of this study was to evaluate whether the addition of synthetic polymers to the vitrification solution affected follicular morphology and development and the expression of Ki-67, Aquaporin 3 (AQP3) and cleaved Caspase-3 proteins in ovarian tissue of the caprine species. Caprine ovaries were fragmented and two fragments were immediately fixed (Fresh Control) for morphological evaluation, while other two were in vitro cultured for 7 days (Cultured Control) and fixed as well. The remaining fragments were distributed in two different vitrification groups: Vitrified and Vitrified/Cultured. Each group was composed of 4 different treatments: 1) Sucrose (SUC); 2) SuperCool X-1000 0.2 % (X-1000); 3) SuperCool Z-1000 0.4 % (Z-1000) or 4) with polyvinylpyrrolidone K-12 0.2 % (PVP). Also, Fresh Control, Cultured Control, SUC and X-1000 were destined to immunohistochemical detection of Ki-67, AQP3 and cleaved Caspase-3 proteins. Morphologically, the treatment with X-1000 showed no significant difference with the Fresh Control group and was superior to the other treatments. After the cleaved caspase-3 analysis, X-1000 showed the lowest percentages of strong immunostaining while Cultured Control showed the highest. Also, a positive correlation was found between the percentages of degenerated follicles and the percentages of strong staining intensity follicles. Regarding the AQP3 analysis, the highest percentages of strong AQP3 staining intensity were found in X-1000. In conclusion, we have demonstrated that the addition of the synthetic polymer SuperCool X-1000 to the vitrification solution improved the current vitrification protocol of caprine ovarian tissue.
AB - The aim of this study was to evaluate whether the addition of synthetic polymers to the vitrification solution affected follicular morphology and development and the expression of Ki-67, Aquaporin 3 (AQP3) and cleaved Caspase-3 proteins in ovarian tissue of the caprine species. Caprine ovaries were fragmented and two fragments were immediately fixed (Fresh Control) for morphological evaluation, while other two were in vitro cultured for 7 days (Cultured Control) and fixed as well. The remaining fragments were distributed in two different vitrification groups: Vitrified and Vitrified/Cultured. Each group was composed of 4 different treatments: 1) Sucrose (SUC); 2) SuperCool X-1000 0.2 % (X-1000); 3) SuperCool Z-1000 0.4 % (Z-1000) or 4) with polyvinylpyrrolidone K-12 0.2 % (PVP). Also, Fresh Control, Cultured Control, SUC and X-1000 were destined to immunohistochemical detection of Ki-67, AQP3 and cleaved Caspase-3 proteins. Morphologically, the treatment with X-1000 showed no significant difference with the Fresh Control group and was superior to the other treatments. After the cleaved caspase-3 analysis, X-1000 showed the lowest percentages of strong immunostaining while Cultured Control showed the highest. Also, a positive correlation was found between the percentages of degenerated follicles and the percentages of strong staining intensity follicles. Regarding the AQP3 analysis, the highest percentages of strong AQP3 staining intensity were found in X-1000. In conclusion, we have demonstrated that the addition of the synthetic polymer SuperCool X-1000 to the vitrification solution improved the current vitrification protocol of caprine ovarian tissue.
KW - In vitro culture
KW - Polyvinylpyrrolidone
KW - SuperCool X-1000
KW - SuperCool Z-1000
KW - Vitrification
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U2 - 10.1016/j.acthis.2019.151484
DO - 10.1016/j.acthis.2019.151484
M3 - Article
C2 - 31902536
AN - SCOPUS:85077284416
SN - 0065-1281
VL - 122
JO - Acta Histochemica
JF - Acta Histochemica
IS - 2
M1 - 151484
ER -