Cryopreservation of caprine ovarian tissue using dimethylsulphoxide and propanediol

Rodrigues, Ana Paula Ribeiro; Amorim, Christiani Andrade; Costa, Sónia; Matos, Maria Helena Tavares de; Santos, Regiane R.; Lucci, Carolina Madeira; Báo, Sônia Nair; Ohashi, Otávio Mitio; Figueiredo, José Ricardo de

doi:10.1016/j.anireprosci.2003.12.003

Cited by 64 publications

(46 citation statements)

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“…Similar changes were reported following exposure of ovarian tissue of sheep [19] and goats [2,3] to 1.5 M concentrations of the same cryoprotectants.…”

Section: Discussionsupporting

confidence: 76%

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Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Wanderley¹,

Luz²,

Faustino³

et al. 2012

Theriogenology

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].You may download, copy and otherwise use the AAM for non-commercial purposes provided that your license is limited by the following restrictions:(1) You may use this AAM for non-commercial purposes only under the terms of the CC-BY-NC-ND license.(2) The integrity of the work and identification of the author, copyright owner, and publisher must be preserved in any copy. Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol AbstractThe objective was to develop an efficient protocol for cryopreservation of agouti (Dasyprocta aguti) ovarian tissue. Agouti ovarian fragments were placed, for 10 min, in a solution containing MEM and fetal bovine serum plus 1.5 M dimethyl sulfoxide (DMSO), ethylene glycol (EG) or propanediol (PROH); some of those fragments were subsequently cryopreserved in a programmable freezer. After exposure and/or thawing, all samples were fixed in Carnoy prior to histological analysis. To evaluate ultrastructure, follicles from the control and all cryopreserved treatments were fixed in Karnovsky and processed for transmission electron microscopy. After exposure and freezing, there was a significant decrease in the percentage of morphologically normal preantral follicles in all treatments when compared to the control (92.67 ± 2.79, mean ± SD). However, there were no significant difference when the exposure and freezing procedures were compared using the same cryoprotectant. Moreover, there was no significant difference among cryoprotectants at the time of exposure (DMSO: 64.7 ± 3.8; EG: 70.7 ± 11.2, PROH: 63.3 ± 8.5) or after freezing (DMSO: 60.6 ± 3.6, EG: 64.0 ± 11.9; PROH: 62.0 ± 6.9). However, only follicles frozen with PROH had normal ultrastructure. In conclusion, preantral follicles enclosed in agouti ovarian tissue were successfully cryopreserved using 1.5 M PROH, with satisfactory maintenance of follicle morphology and ultrastructure.

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“…Similar changes were reported following exposure of ovarian tissue of sheep [19] and goats [2,3] to 1.5 M concentrations of the same cryoprotectants.…”

Section: Discussionsupporting

confidence: 76%

“…animals, including sheep [1], goats [2,3], primates [4], rodents [5], and humans [6]. Satisfactory results have been reported, especially when a 1.5 M concentration was used in slow-freezing protocols.…”

Section: Introductionmentioning

confidence: 99%

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Wanderley¹,

Luz²,

Faustino³

et al. 2012

Theriogenology

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“…Luna & Munhoz (2008) successfully vitrified bovine ovarian tissue fragments, using dimethylsulfoxide (2.6 M). In caprine preantral follicles cryopreserved with dimethylsulfoxide, remained good conditions the granulosa and nucleus cells (Rodrigues et al 2004). Castro et al (2011) used dimethylsulfoxide as intracellular cryoprotectant agent and demonstrated its effectiveness in preserving caprine ovarian tissue.…”

Section: Experiments Imentioning

confidence: 99%

Vitrification of bovine preantral follicles with dimethylsulfoxide and sucrose plus α-tocopherol

et al. 2016

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RESUMO.-[Vitrificação de folículos pré-antrais bovinos com dimetilxulfoxido e sacarose adicionado de α-tocopherol.] Os objetivos deste estudo foram avaliar a vitrificação de folículos pré-antrais bovinos com dimetilsulfóxido (D) e sacarose (S) adicionando α-tocoferol 5mmol/L (T5) ou 10mmol/L (T10) e, avaliar o aquecimento com meio essencial mínimo (m) com ou sem sacarose (s). Ovários de fêmeas bovinas foram coletados de abatedouro, para o experimento I (n= 66) e II (n= 51). No laboratório fragmentos ovarianos foram distribuídos aleatoriamente para o controle fresco e 8 tratamentos de vitrificação (Controle e Dm; Dms, a DSm; DSms; DST5m; DST5ms; DST10m; DST10ms). Os fragmentos ovarianos foram colocados na solução de vitrificação (5 min) e imersos em nitrogênio líquido (-196°C The objective of this study was to evaluate the vitrification of bovine preantral follicles with dimethylsulfoxide (D) and sucrose (S) plus α-tocopherol 5mmol/L (T5) or 10mmol/L (T10) and, evaluate the thawed with minimal essential medium (m) with or without sucrose (s). Ovaries of cows were collected from slaughterhouse for the experiment I (n=66) and II (n=51). In the laboratory ovarian fragments were randomly assigned either to fresh control and 8 vitrification treatments (Controle and Dm; Dms, DSm; DSms; DST5m; DST5ms; DST10m; DST10ms). Ovarian fragments were placed in vitrification solution (5 min) and immersed in liquid nitrogen (-196°C), after a week, the fragments were thawed and analyzed. In the experiments I, preantral follicles were morphologically observed for histological evaluation, (normal; degenerated and developing of stage). In the experiment II, preantral follicles were mechanically isolated from ovarian tissue and examined with trypan blue, where dead and live corresponded to stained or non-stained. The treatments DSm, DSms and DST10m were effective in preserving the morphology in situ. However, the viability of isolated preantral follicles after vitrification remained high only in treatment DST10m. Thus, DST10m preserves survival rates and morphological integrity during vitrification of bovine preantral follicles. tamento DST10m. Assim, DST10m preservou as taxas de sobrevivência e integridade morfológica durante a vitrificação de folículos pré-antrais bovinos.

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“…Most of the preservation protocols for ovarian tissue have been performed in presence of permeable cryoprotective substances such as glycerol (GLY), ethylene glycol (EG), propanediol (PROH) and dimethyl sulfoxide (DMSO), in concentrations ranging from 1.5 to 3.0 M. Besides their cryoprotective effect, these substances can also be toxic to cells, mainly during exposure period (Rodrigues et al, 2004a, Rodrigues et al, 2004band Santos et al, 2007b. Therefore, attention must be given to ovary transport before cryopreservation procedures.…”

Section: Introductionmentioning

confidence: 99%

“…The use of histology alone is insufficient to assess the freezing-thawing process as morphological analysis, and is often not correlated to the viability or developmental competence of the follicles (Santos et al, 2006a andSantos et al, 2007a). Transmission electronic microscopy (TEM) is an important tool to assess the cell quality after freezing-thawing, because cryopreservation often induces loss of cell contents (Rodrigues et al, 2004a) and loss of mitochondrial activity (Santos et al, 2006b). In studies using ovine embryos as model, Cocero et al (2002) showed damage in organelles, at the ultra-structural level, from apparently normal cells.…”

Section: Introductionmentioning

confidence: 99%

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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Cryopreservation of caprine ovarian tissue using dimethylsulphoxide and propanediol

Cited by 64 publications

References 50 publications

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Vitrification of bovine preantral follicles with dimethylsulfoxide and sucrose plus α-tocopherol

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

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