Morphokinetic changes in vitrified and non-vitrified in vitro-derived ovine embryos

Fryc, Karolina; Nowak, Agnieszka; Kij, Barbara; Kochan, Joanna; Bartlewski, Pawel M.; Murawski, M.

doi:10.1016/j.theriogenology.2022.04.027

Cited by 3 publications

(3 citation statements)

References 24 publications

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Section: Discussionmentioning

confidence: 73%

“…We found that increased numbers of freeze-thaw cycles is associated with a decline in cryosurvival and a higher number of non-viable embryos, indicating deteriorating embryo quality. Supporting this, recent literature in an ovine model has shown that vitrified embryos are more susceptible to cellular apoptosis and exhibit lower cryosurvival rates compared to their non-vitrified counterparts 14 . Furthermore, vitrification-induced cellular apoptosis of blastocyst-stage embryos is known to be associated with mitochondrial dysfunction, compromising the developmental potential of bovine and porcine embryos [23][24] .…”

Section: Discussionmentioning

confidence: 73%

“…While many studies have showed that vitrification, an ultra-rapid cryopreservation procedure, has comparable pregnancy rates and postnatal outcomes compared to fresh embryo transfers [7][8][9][10][11] , there are contradictory studies that showed detrimental impact on clinical pregnancy rate and live birth rate [12][13] . Additionally, a recent study in an ovine model showed that cryopreserved embryos are prone to apoptosis and have lower cryosurvival rates compared to non-cryopreserved embryos 14 . However, the impact of cryopreservation on the developmental and implantation potential of embryo remains unclear.…”

Section: Introductionmentioning

confidence: 99%

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Impact of repeated cryopreservation on embryo health and implantation potential

Li,

Chow

et al. 2023

Preprint

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In IVF clinics, preimplantation genetic testing (PGT) is a common practice that involves a biopsy and cryopreservation of embryos for genetic evaluation. When testing fails or is required for already cryopreserved embryos, multiple freeze-thaw cycles occur. Though known to impact live birth rates, the exclusive influence of cryopreservation has not been elucidated. Here, we evaluate the effect of repeated cryopreservation on embryo health and implantation potential. Blastocyst-stage murine embryos were subjected to one, two or three freeze-thaw cycles with fresh embryos serving as a control. Outcomes assessed included post-thaw survival rate, allocation of cells to the inner cell mass (ICM) vs. trophectoderm cell lineages, implantation potential and offspring health. Post-thawing, embryos that were subjected to three freeze-thaw cycles had a significantly lower survival rates compared to embryos that had undergone one cycle (P<0.001). Additionally, the number of ICM cells was significantly reduced in embryos subjected to two or three freeze-thaw cycles compared to fresh or single-cycle embryos (P<0.001). No statistically significant differences were found for pregnancy rate, number of implantations, viable fetuses or resorption sites between treatment groups. We did however, find a non-significant yet interesting trend: three freeze-thaw cycles were associated with a 20% decrease in viable fetuses and a 20% increase in resorption sites compared to one freeze-thaw cycle group. These findings demonstrate that repeated cryopreservation adversely affects embryo health and may decrease implantation potential. Consequently, caution is advised regarding the repeated application of cryopreservation in IVF clinics, underscoring the need for further research to optimise cryopreservation protocols.

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Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 73%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Impact of repeated cryopreservation on embryo health and implantation potential

Li,

Chow

et al. 2023

Preprint

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Morphokinetic changes and apoptotic cell death in vitrified and non-vitrified in vitro-produced ovine embryos

Fryc

Nowak

Kij

et al. 2023

Reproductive Biology

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Analysis of cryoresistance of cumulus of Ovis Arias oocytes after intra- or extra-ovarian vitrification

Kuzmina

Sokolova

2022

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The aim of this study was to identify the effects of highly dispersed silica nanoparticles (HDSns) on the cryoresistance of Ovis Aries cumulus of oocytes in conditions of intra-(IOV) or extra-(EOV) ovarian vitrification.Materials and methods. Cumulus-oocyte complexes (COC) intended for EOV were treated with solutions of cryoprotectants (CPA) prepared on TC-199 with 10% FBS (fetal bovine serum): CPA-1-0.7 M dimethyl sulfoxide (DMSO) + 0.9 M ethylene glycol (EG);CPA-2-1.4 M DMSO + 1.8 M EG; CPA -3- 2.8 M DMSO + 3.6 M EG + 0.65 M trehalose. The COC was gradually exposed for 30 seconds in CPA-1, 30 seconds in CPA-2, and 20 seconds in CPA-3. At the IOV, the ovaries of the animals were divided into fragments and placed in sterile gauze containers in CPAs prepared on the basis of Dulbecco (PBS) with 20% FBS: CPA-1 - 7.5% EG + 7.5% DMSO (25 min), then in CPA-2 - 15% EG+15% DMSO + 0.5 M sucrose (15 min). 0.001% HDSns (A.A. Chuiko ICP of the National Academy of Sciences of Ukraine) were added to the CPAs and media for thawing of the experimental groups. The straws with the biomaterial were stored in liquid nitrogen for at least 24 hours. The COCs were removed from the straws after thawing and placed successively at 3 min: in a 0.25 M solution of trehalose in TC-199 with 10% FBS at 37°C, then in 0.19 M, and finally in 0.125 M trehalose. COCs from the thawed fragments were sequentially treated with 0.5 M (1 min) and 0.25 M (5 min) solutions of sucrose in PBS with 20% FBS. The final washing of all COCs was carried out in TC-199 with 10% FBS.Results. The results of morphological (the character of oocyte-cumulus communications) and cytological monitoring (the status of cumulus cells chromatin) revealed a positive effect of 0.001% HDSns on the parameters of cryoresistance of COC both at EOV and IOV. The proportion of cells with compact cumulus in the group of EOV of gametes with HDSns significantly exceeded that in their absence (69 % vs. 51 %, P<0.05). A similar trend was shown in IOV of oocytes (35 % vs. 20 %, P<0.05). The level of apoptotic (TUNEL-test) cells in the group of EOV (44 % vs. 32 %, P<0.05) and IOV (72 % vs. 51 %, P<0.001) of gametes sharply decreased when HDSns were added to cryoprotective media, and the proportion of pyknotic cells decreased during EOV (from 31 % to 21 %, P<0.05) and IOV (from 58 % to 41 %, P<0.05). In general, the obtained results indicate the expediency of using 0.001 % HDSns in the protocols of IOV and EOV of female gametes OVIS ARIAS.

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Morphokinetic changes in vitrified and non-vitrified in vitro-derived ovine embryos

Cited by 3 publications

References 24 publications

Impact of repeated cryopreservation on embryo health and implantation potential

Impact of repeated cryopreservation on embryo health and implantation potential

Morphokinetic changes and apoptotic cell death in vitrified and non-vitrified in vitro-produced ovine embryos

Analysis of cryoresistance of cumulus of Ovis Arias oocytes after intra- or extra-ovarian vitrification

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