Abstract
The objective of this study was to evaluate the effect of ovarian fragments vitrification in the morphometry, number of granulosa cells and nuclear chromatin of preantral ovarian follicles of bovines. Ovarian was obtained at a local slaughterhouse and cortex fragments were removed. One third wereimmediately fixed and processed for histology (T0 - control). The remaining fragments were divided in two treatments, TI - toxicity test and TII - vitrification. In TI the fragments were exposed to only cryoprotectants and fixed in Carnoy and in TII were utilized DMSO and ethylene glycol supplemented with sucrose. The results indicated that the primordial follicles when vitrified or exposed to cryprotectants did not show variation in morphometry and amount of cells of the granulosa (p > 0.05), exception to primary follicles (p<0.05) when compared to T0. The analysis of the nuclear chromatin of follicles vitrified or cryprotectants exposed showed significant difference (p<0.05) when compared to T0 with the control treatment, particularly in TII. In conclusion the vitrification technique affected negatively the morphology of bovine follicles enclosed in ovarian tissue.
Highlights
The objective of this study was to evaluate the effect of ovarian fragments vitrification in the morphometry, number of granulosa cells and nuclear chromatin of preantral ovarian follicles of bovines
In TI the fragments were exposed to only cryoprotectants and fixed in Carnoy and in TII were utilized DMSO and ethylene glycol supplemented with sucrose
The results indicated that the primordial follicles when vitrified or exposed to cryprotectants did not show variation in morphometry and amount of cells of the granulosa (p>0.05), exception to primary follicles (p
Summary
Obtiveram-se doze ovários de vacas em abatedouro, os quais foram transportados em bolsas plásticas contendo solução salina de NaCl a 0,9%, em temperatura ambiente, até o laboratório. Dos doze ovários se retiraram 36 fragmentos com aproximadamente 3 mm x 1 mm cada um, que foram distribuídos aleatoriamente para três tratamentos: T0 – controle, fixados imediatamente; TI – expostos aos crioprotetores e TII – expostos aos crioprotetores e vitrificados. Os fragmentos no TI destinados apenas ao teste de toxicidade foram expostos nas mesmas soluções crioprotetoras e tempos de exposições como no TII; após a exposição, os fragmentos foram fixados em Carnoy durante 12 h e submetidos a rotina histológica clássica. Para a análise folicular usou-se o microscópio óptico em aumento de 400 x; inicialmente, realizou-se a classificação dos folículos pré-antrais de acordo com o estádio de desenvolvimento como: primordiais: uma camada de células da granulosa de forma pavimentosa ou pavimentosacubóide ao redor do ovócito; primários: uma única camada de células da granulosa de forma cubóidal, e pequenos secundários: duas ou mais camadas de células da granulosa de forma cúbica. Morfometria, células da granulosa e cromatina nuclear de folículos pré-antrais em associação ou nãocom a vitrificação de tecido ovariano bovino 373
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