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

Abstract

In contributing to the conservation of wild rodents, the aim of this study was to evaluate the use of distinct cryoprotectants, separately or in combination, for solid surface vitrification (SSV) of red‐rumped agouti ovarian tissue. Ovarian cortex from nine females was recovered and fragmented. Fresh fragments (control) were used to analyse the pre‐antral follicle (PF) morphology using a histologic procedure, viability using the Trypan blue test, cell proliferation by counting the argyrophilic nucleolar organizing regions (Ag‐NORs technique) and DNA integrity using the TUNEL assay. The remaining fragments were vitrified using SSV method with 3 M or 6 M ethylene glycol (EG) or dimethyl sulfoxide (DMSO), or in combination (3 M EG/3 M DMSO), and further evaluated as reported for the fresh samples. All cryoprotectants were effective at preserving PFs morphology compared to the control group (80.7 ± 5.21%), except 6 M EG and 3 M DMSO that provoked a significant (p < .05) decrease on the values of morphologically normal primary (60.0 ± 19.0%) and primordial (44 ± 4.5%) follicles, respectively. Regarding viability, all cryoprotectants provided values similar to that verified for the control group (79.0%), but a significant decrease (p < .05) was observed with EG/DMSO combination (59%). Using Ag‐NORs technique, the highest (p < .05) cell proliferative capacity was detected when using EG at each tested concentration. The TUNEL proved the preservation of DNA integrity regardless of the cryoprotectant. In summary, we suggest the use of 3 M EG for the solid surface vitrification of red‐rumped agouti ovarian tissue.

Reproductive Technologies Used in Female Neo-Tropical Hystricomorphic Rodents

“…Analysis of the results indicated that PROH was the best cryoprotectant agent of the three evaluated and thus was recommended. The use of DMSO and EG for solid-surface vitrification (SSV) was also proven effective in the preservation of ovarian tissue, with 3 M EG being the most effective [ 16 ].…”

“…Agoutis ( Dasyprocta spp.) are medium-sized rodents that weigh between 2–3.5 kg [ 13 , 14 , 15 , 16 , 17 ]. Studies defining the age of puberty in females are lacking, however, Guimarães et al [ 18 ] showed that, under the male effect, puberty in D. prymnolopha was attained at nine months.…”

In vitro culture of red-rumped agouti preantral follicles enclosed in fresh and vitrified ovarian tissues using TCM199 plus different pFSH concentrations

“…From the 13 catalogued agouti species, only three remain presenting a stable population, including the Dasyprocta leporina , the red-rumped agouti, that inhabits Brazilian Caatinga ( Emmons and Reid, 2016 ). At this sense, various efforts related to ex situ conservation strategies, as the biobank formation ( Castelo et al, 2015 ; Praxedes et al, 2020 ; Rodrigues et al, 2021 ), have been conducted to help on the efforts to maintain the stability of free-living D. leporina populations and provide technologies for its captive sustainable breeding ( Lall et al, 2020 ).…”

Biobanking and use of gonadal tissues - a promising strategy for conserving wildlife from the Caatinga biome

“…Protocols to create BRB from ovarian and testicular tissues have been developed in different wild species ( Campos et al, 2019b , Praxedes et al, 2020 , Silva et al, 2021a , b , 2022 ). In general, the efficiency of BRB of preantral follicles (PAFs) enclosed in ovarian tissues or germ cells enclosed in testicular tissues depend on the technical conditions involved in the recovery, cryopreservation, in vivo or in vitro culture and characterization of these tissues, which are all species-specific ( Campos et al, 2019b , Silva et al, 2020 ).…”