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

Abstract

].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.

Clinically applied procedures for human ovarian tissue cryopreservation result in different levels of efficacy and efficiency

“…In accordance with the results of the current study, a better follicle morphology and a higher percentage of living follicles were found after cryopreservation using DMSO when compared to ethylene glycol for human, sheep, or goat ovarian tissue [27–30]. However, DMSO did not improve the preservation of the follicle survival and ultrastructure when compared to ethylene glycol in bovine or agouti tissue [31, 32]. …”

Monitoramento do ciclo estral de cutias (Dasyprocta leporina Lichtenstein, 1823) através de citologia esfoliativa vaginal e ultrassonografia

“…Nos últimos anos, diversos conhecimentos têm sido gerados no tocante à biologia reprodutiva das cutias, como a caracterização morfológica dos ovários (Almeida et al 2003), tubas uterinas (Fortes et al 2005), placentação (Rodrigues et al 2003) e criopreservação de folículos pré-antrais (Wanderley et al 2012) em D. leporina, bem como o diagnóstico de gestação por ultrassonografia em D. prymnolopha (Sousa et al 2012). Das 11 diferentes espécies de cutias catalogadas, genericamente conhecidas como D. aguti, apenas sete habitam o território brasileiro (Woods & Kilpatrick 2005), no entanto, diferentes aspectos reprodutivos entre as diferentes espécies permanecem não elucidados.…”

Comparison of different intracellular cryoprotectants on the solid surface vitrification of red‐rumped agouti (Dasyprocta Leporina Lichtenstein, 1823) ovarian tissue

“…We provide unprecedent data regarding the use of different cryoprotectants for the solid surface vitrification (SSV) of red-rumped agouti ovarian tissue. We demonstrated that it is possible to preserve up to 70% PFs morphology after SSV, especially with the use of 3 M EG with a better result than that previously reported for the same species using the slow-freezing process (~60%) (Wanderley et al, 2012). One of the advantages in considering SSV is the use of a metal that acts as a heat conductor to allow for the rapid cooling of the samples (Gupta, Uhm, & Lee, 2007).…”