Cryopreservation of Preantral Follicles

In mammals, the total number of female germ cells is already established by the time of birth, meaning that no mitosis will take place in oogonias thereafter. Their cryostorage, therefore, depends on ovarian tissue manipulation. As an alternative to mature oocyte cryopreservation, the maintenance of inactive preantral follicles is a remarkable option because (i) their availability in the ovary is greater; (ii) as inactive and small structures, they show less sensitivity to cryoinjury and the toxic effects of cryoprotectants; and (iii) they are present in the gonads at all ages, allowing their retrieval from prepubertal individuals or even immediately postmortem. Nevertheless, some difficulties remain regarding their in vitro activation and development to the ovulatory stage. For this reason, the best option for their total development is transplantation back to the donor or between species, promoting follicle activation and development. This technique has proved its efficiency and led to several live births in both animals and humans. Since each species has its own particularities in terms of ovarian tissue composition, a number of protocols have been documented, which may be used for either isolated or in situ preantral follicles

Avaliação morfológica dos testículos de ovinos jovens Santa Inês submetidos a diferentes regimes de suplementação alimentar e vermifugação

ABSTRACT: The present study investigated the effect of protein supplementation and parasite load in relation to testicular morphology, in young Santa Inês male sheep. Twentyfour male lambs, non-castrated, were distributed in four treatments: HPd (dewormed animals + diet with high protein), HPn (animals non dewormed + diet with high protein), LPd (dewormed animals + diet with low protein), LPn (animals not dewormed + diet with low protein) during eight months. At the end of the experiment histological cuts of testicles were taken and cell count analyzed by computer and the program IMAGE PRO-PLUS was used for evaluation of measurements of the seminiferous tubules. The following results were found: LPn presented larger number of spermatogonias, but a reduced number of the other cells parameters (Sertoli cells, spermatocytes, spermatids and spermatozoa) and smaller diameter and circumference of the seminiferous tubules and shorter seminiferous epithelium when compared to other treatments. There was a significant correlation among tubules diameter of the epithelium height and tubules circumference. Correlations between lumen diameter and lumen circumference, as well as epithelium height and tubules circumference also presented high and positive correlations (r>0.70). When the external testis parameters were compared with internal parameters no significant correlation was observed (r0,70); comparando-se os parâmetros testiculares externos com os internos, observa-se que não houve correlação significativa (P<0,50). Conclui-se que a alta proteína influencia positivamente a espermatogênese, e a não-vermifugação associada a uma dieta pobre em proteínas pioram substancialmente a maioria dos parâmetros reprodutivos de carneiros jovens Santa Inês

Mechanical isolation of caprine preantral follicles at different reproductive stages and ovarian situation

O presente trabalho descreve os efeitos do estágio reprodutivo (animais impúberes, adultos não-gestantes e gestantes), posição anatômica do ovário (direito e esquerdo) presença de corpo lúteo funcional, peso do ovário e número de folículos antrais superficiais (FAS) sobre o número de folículos pré-antrais (FPA) isolados mecanicamente. Os FPA isolados foram divididos em 3 classes a saber: primordiais, primários e secundários. O número médio (± EP) de FPA isolados por ovário foi de 1.324 ± 193; 866 + 170 e 779 ± 139, respectivamente para ovários oriundos de animais impúberes, não-gestantes e gestantes, sendo observada diferença significativa somente entre animais impúberes e gestantes. A posição anatômica do ovário (exceto para os animais não-gestantes) e a presença ou ausência de corpo lúteo não exerceram nenhum efeito sobre o número e distribuição dos FPA isolados. De um modo geral, o número de folículos primários e secundários foi negativamente correlacionado com o peso ovariano. Já o número de folículos primordiais e secundários foi, respectivamente, positiva e negativamente correlacionado com o número de FAS. Analisando as categorias de animais isoladamente, observou-se que, nos animais impúberes, o número de folículos primordiais foi positivamente correlacionado com o número de FAZ, enquanto que o número de folículos primários e secundários foi negativamente correlacionado com o número de FAS. No tocante à categoria de animais gestantes, o número de folículos primários foi negativamente correlacionado com o peso ovariano. Dependendo da categoria de ovários estudada, foram observadas associações significativas entre o número de folículos primordiais, primários e secundários. _________________________________________________________________________________ ABSTRACTThe aïm of this study was to describe the effects of reproductive stages (pre-pubertal, non-pregnant and pregnant adult animals), ovarian anatomic position (left or right), presence of functional corpus luteum, ovarian weight and number of superficial antral follicles on the number of preantral follicles mechanically isolated. The mean number (mean ± SEM) of isolated preantral follicles per ovary was 1,324 ± 193, 866 ±170 and 779 ± 139, respectively for prepubertal, non-pregnant and pregnant adult animals. Anatomic position (except for nonpregnant animals) and the presence or absence of corpus luteum dia nof affect the number and distribution of isolated preantral follicles. In general, the number of primary and secondary follicles was negatively correlated with the ovarian weight. The numbers of primordial and secondary follicles were, respectively, positively and negatively correlated with the number of superficial antral follicles. Analyzing animal categories separately, it was observed that in prepurbetal animals, the number of primordial follicles was positively correlated with the number of superficial antral follicles. However the numbers of primary and secondary follicles were negatively correlated with the number of superficial antral follicles. In the category of pregnant animals, the number of primary follicles was negatively correlated with the ovarian weight. According to ovary category analyzed, were observed significant correlation between the number of primordial, primary and secondary isolated follicles

Study of preantral follicle population in situ and after mechanical isolation from caprine ovaries at different reproductive stages

The purposes of this study were to estimate the population of caprine preantral follicles, and to evaluate quantitatively and qualitatively the efficiency of a specific mechanical method for the isolation of preantral follicles from mixed breed goats at different reproductive stages. On average, 37,646±4277 preantral follicles were present in goat ovaries, and 13,631±2399 preantral follicles were obtained after isolation. The number of preantral follicles isolated or in situ was not significantly affected by the reproductive stage. The mean recovery rate per ovary ([number of isolated follicles/number of in situ follicles]×100) of isolated follicles was 36.2%. The distribution of follicles in situ was 67.8% primordial, 25.8% primary and 6.4% secondary; the respective distribution after isolation was 93.8%, 5.2% and 1.0%. In this study, many polyovular follicles were also observed, mainly in prepubertal goat ovaries. Histological analysis showed that few preantral follicles were atretic in situ (4.83%±0.35) or after the isolation procedure (4.67%±0.65) in the three reproductive stages. The percentage of atretic follicles was not affected either by the mechanical method or by the reproductive stage. It is concluded that a large number of preantral follicles can be successfully isolated mechanically, with a high recovery rate and a low rate of follicular atresia, irrespective of the reproductive stage of the caprine female

Mejoras y perspectivas en criopreservación de tejido ovárico y testicular

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