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Reprod Domest Anim ; 54(3): 480-485, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30444528


This study aimed to examine the in vitro culture of secondary preantral follicles, using reused ovaries, to compare both the 2D and 3D methods of in vitro culture of preantral follicles, and the system of medium replacement. Twenty-five pairs of ovaries from mixed-breed goats were used for the experiment. Follicular puncture of antral follicles was performed for in vitro production. After this procedure, the secondary preantral follicles were submitted to a microdissection procedure. The isolated preantral follicles were randomly divided into three treatments: (a) Two-dimensional culture with partial replacement of medium during culture (2D PR), (b) Three-dimensional culture with addition of medium during culture (3D AD) and (c) Three-dimensional culture with partial replacement of medium (3D PR). The culture period was 18 days. All treatments at the end of the in vitro culture period (18 days) presented a follicular survival rate which ranged from 59% to 70%, demonstrating that it was possible to perform an experiment with preantral follicles using ovaries that had previously been used in another reproductive biotechnique. The 3D AD treatment showed a survival percentage and follicular diameter higher than the 2D PR treatment, however, it did not differ from the 3D PR treatment. In conclusion, experiments employing the use of preantral follicles can be performed with success after the ovaries have been used for experiments with antral follicles. Moreover, the three-dimensional system with the addition of medium is recommended for in vitro culture of preantral follicles, since this system is more practical and financially feasible.

Folículo Ovariano/crescimento & desenvolvimento , Ovário/fisiologia , Técnicas de Cultura de Tecidos/veterinária , Animais , Meios de Cultura , Feminino , Hormônio Foliculoestimulante/farmacologia , Cabras , Oócitos/metabolismo , Reprodução , Técnicas de Cultura de Tecidos/métodos
Braz. arch. biol. technol ; 57(2): 200-208, Mar.-Apr. 2014. ilus, graf, tab
Artigo em Inglês | LILACS-Express | LILACS | ID: lil-705746


The aim of the present study was to determine the role of GDF-9 and/or FSH on the growth and mRNA expression for FSH-R, GDF-9, and BMPs in goat secondary follicles after culture in vitro. Goat secondary follicles (~200µm) were isolated and cultured for six days in minimum essential medium (MEM) supplemented with GDF-9 (200 ng/mL), FSH (50 ng/mL) or both. At the beginning and end of culture, the follicular diameter was evaluated and compared. The levels of mRNA for GDF-9, FSH-R and BMPs -2, -4, -6, -7 and -15 in cultured follicles were quantified by real time PCR. The results showed that a significant increase of follicle diameter after six days when compared to day 0, but the presence of GDF-9 and FSH did not influence the follicular growth in comparison with those cultured in MEM. Real time PCR showed that GDF-9 down-regulated the levels of mRNA for BMPs -2 and -15, while FSH either alone or in combination with GDF-9 did not affect the expression of GDF-9, FSH-R and BMPs. In conclusion, GDF-9 reduced the expression of BMP-2 and -15 in caprine preantral follicles after their culture, but FSH either alone or in association with GDF-9 did not control the expression of GDF-9, FSH-R and BMPs.