Conservação de folículos pré-antrais bovinos em solução salina 0,9% ou TCM 199

Celestino, J.J.H.; Santos, Regiane R.; Martins, F.S.; Matos, Maria Helena Tavares de; Figueiredo, J.R.; Costa, Sónia; Silva, J.R.V.; Rodrigues, Ana Paula Ribeiro

doi:10.1590/s0102-09352007000300007

Cited by 9 publications

(3 citation statements)

References 23 publications

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“…Morphological assessment of follicular integrity has been largely used to evaluate the effectiveness of various treatments (media, temperature and time of preservation) to which ovarian follicles are submitted (Matos et al , 2004; Celestino et al , 2007). However, additional measurements such as in vitro follicular development may be used as a reliable evaluation of follicles and/or oocyte viability (Abd-Allah, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Techniques for short-term storage of ovaries were developed for several species, such as ovine (Matos et al , 2004), caprine (Silva et al , 2000; Chaves et al , 2008), bovine (Celestino et al , 2007), swine (Wongsrikeao et al , 2005) and equine (Love et al , 2003). Most of these studies evaluated only the morphological features of preantral follicles through histological analysis, after storage of ovarian tissue in different temperatures, periods and media (Matos et al , 2004; Celestino et al , 2007). However, additional measurements of follicular viability and developmental competence, such as in vitro preantral follicle culture, were performed only in caprine species (Chaves et al , 2008).…”

Section: Introductionmentioning

confidence: 99%

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Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Gonçalves

Cavalcante

Gouveia

et al. 2015

Zygote

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The aim of this study was to investigate the effect of ovarian tissue transportation conditions (medium and period of time) on the morphology, apoptosis and development of ovine preantral follicles cultured in vitro. Each ovarian pair was cut into nine slices, with one fragment being fixed immediately (fresh control). The remaining fragments were placed individually in cryotubes containing conservation medium (minimal essential medium (MEM) without supplementation or MEM+ - with supplementation) and stored at 35ºC for 6 or 12 h without (non-cultured) or with subsequent culture for 5 days. Then, the fragments were processed for histological and terminal deoxynucleotidyl transferase (TdT) mediated dUTP nick-end labelling (TUNEL) examination. Preservation of ovarian slices in MEM or MEM+ (non-cultured) resulted in similar percentages of normal follicles when compared with the fresh control. Nevertheless, compared with the fresh control, a decrease in the percentage of normal follicles was observed in tissues cultured for 5 days. Only for tissues preserved in supplemented medium (MEM+) for 6 h, the percentage of TUNEL positive cells was similar between non-cultured tissues and tissues cultured for 5 days. Follicular activation and growth (follicular and oocyte diameter) were higher in cultured tissues than in fresh control or non-cultured tissues, except those from fragments preserved for 6 h in MEM and then cultured for 5 days in which no growth was observed. In conclusion, ovine ovarian tissue was successfully preserved in supplemented medium (MEM+) at a temperature close to physiological values (35°C) for up to 6 h without affecting apoptosis in the ovarian follicles and their ability to develop in vitro.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Gonçalves

Cavalcante

Gouveia

et al. 2015

Zygote

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“…Deve ser ressaltado que o transporte de ovários com a presença da bursa, como realizado no presente trabalho, é um fator que pode promover a proteção dos oócitos conservando--se a sua temperatura resfriada. Celestino et al (2007) verificaram que o metabolismo fisiológico (39°C) associado à baixa tensão de oxigênio in vitro provoca a degeneração folicular. Essas afirmações podem explicar os valores encontrados no presente trabalho das altas taxas de degeneração oocitária observadas em oócitos obtidos nas fases de anestro e diestro do ciclo estral.…”

Section: Discussionunclassified

Influência da temperatura de transporte de ovários na maturação in vitro de oócitos caninos coletados em diferentes estágios do ciclo estral

Pereira

Bersano

Lopes

2015

Braz. J. Vet. Res. Anim. Sci.

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ResumoFoi avaliada a influência do ciclo estral e temperatura de transporte de ovários na maturação in vitro de oócitos caninos. As cadelas foram categorizadas em dois grupos baseados no estágio do ciclo estral -anestro ou diestro. Um ovário por par coletado foi transportado em solução fisiológica 0,9% à 4°C enquanto o outro foi transportado a 37°C. Então, os ovários foram seccionados em PBS para a liberação dos complexos cumulus oócito (COCs). Um total de 345 COCs (n = 186 oócitos obtidos de cadelas em anestro e 159 em diestro) foi cultivado em TCM 199 suplementado com HEPES, piruvato de sódio, cisteína, hormônio folículo estimulante (FSH), gonadotrofina coriônica humana (hCG), estrógeno (E 2 ) e fator de crescimento epidermal (EGF). Após 72h de maturação, os COCs foram desnudados, fixados e corados para avaliação da maturação nuclear. O teste de Fisher foi utilizado para avaliar as diferenças entre os grupos. O nível de significância adotado foi de 0,05. Os oócitos obtidos de cadelas em diestro transportados a 4°C apresentaram maior frequência de oócitos no estágio de metáfase II (21,1%) que os mantidos na temperatura de 37°C (p < 0,01). De forma similar, houve maior frequência de oócitos nos estágios de metáfase II (11,2%) nos ovários obtidos de cadelas em anestro e transportados a 4°C que nos ovários mantidos a 37°C (p < 0,05). Concluiu-se que a temperatura de transporte influencia os resultados de viabilidade oocitária canina e a maturação in vitro, independentemente do estágio reprodutivo da fêmea. Palavras-chave:Maturação in vitro. Temperatura. Anestro. Diestro. Cadela. AbstractThis study evaluated the influence of estrous cycle stage and transport temperature of ovaries on in vitro maturation of canine oocytes. The bitches were categorized into two groups based on stage of estrus cycle: diestrus or anestrus. One ovary of each pair collected was transported in saline solution at 4°C while the other was transported at 37°C. Thus, ovarian tissue was sliced in PBS to release cumulus oocyte complexes (COCs). A total of 345 COCs (n = 186 oocytes from ovaries of bitches in anestrus and 159 in diestrus) were cultivated in TCM 199 supplemented with HEPES, sodium pyruvate, cysteine, follicle stimulating hormone (FSH), human chorionic gonadotropin (hCG), estrogen (E 2 ) and epidermal growth factor (EGF). After 72h of maturation, the COCs were denuded, fixed and stained to assess nuclear maturation. The Fisher test was applied to examine the differences between the groups. The significance level adopted was 0.05. The oocytes obtained from the ovaries from bitches in diestrus transported at 4°C shown increased frequency of oocytes in metaphase II stage than those maintained at 37°C (p < 0.01). Similarly, there was increased frequency of oocytes in metaphase II (11.1%) stage from the ovaries of bitches in anestrus and transported at 4°C, than those maintained at 37°C (p < 0.05). It was concluded that transport temperature influences the results of canine oocyte viability and in vitro maturation, regardless of reproduc...

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Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue

Celestino¹,

Santos

Lopes³

et al. 2008

Animal Reproduction Science

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Bovine preantral follicles within ovarian fragments were exposed and cryopreserved in absence or presence of 1.5 M glycerol (GLY), ethylene glycol (EG), propanediol (PROH) or dimethyl sulfoxide (DMSO), undergoing a previous cooling at 20 °C for 1 h (protocol 1) or at 4 °C for 24 h (protocol 2) in 0.9% saline solution. At the end of each treatment, preantral follicles were classified as non-viable/viable when they were stained/not stained with trypan blue, respectively. To confirm viability staining, ultra-structure of the follicles was evaluated by transmission electronic microscopy (TEM). Data were compared by Chi-square test (P < 0.05). The storage of the ovaries at 20 °C for 1 h (78%) and 4 °C for 24 h (80%) did not reduce significantly the percentage of viable preantral follicles when compared to the control (75%). Similar results were obtained when ovarian fragments, respectively, for protocols 1 and 2, were exposed to MEM (78 and 77%), 1.5 M EG (78 and 71%), as well as frozen in 1.5 M EG (74 and 77%). Percentages of viable follicles in control were similar to those observed after exposure (75%) and freezing (76%) in presence of 1.5 M DMSO only when protocol 1 was used. The increase of the concentration from 1.5 to 3.0 M, for all cryoprotectants, reduced significantly the percentage of viable preantral follicles after freezing. Ultra-structural analysis has confirmed trypan blue results, showing that not only basement membrane, but also organelles, were intact in viable preantral follicles. In conclusion, ovarian tissue cooling at 4 °C for 24 h before cryopreservation (protocol 2) does not affect the viability of bovine preantral follicles when 1.5 M EG is present in the cryopreservation medium.

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Conservação de folículos pré-antrais bovinos em solução salina 0,9% ou TCM 199

Cited by 9 publications

References 23 publications

Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Lower apoptosis rate in ovine preantral follicles from ovaries stored in supplemented preservation media

Influência da temperatura de transporte de ovários na maturação in vitro de oócitos caninos coletados em diferentes estágios do ciclo estral

Preservation of bovine preantral follicle viability and ultra-structure after cooling and freezing of ovarian tissue

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