Tolerance of lamb and mouse oocytes to cryoprotectants during vitrification

Abstract

SummaryMouse and lamb oocytes were vitrified with, or exposed to, different cryoprotectants and evaluated for their effects on their survival and developmental competence after in vitro fertilization (IVF) and activation treatments. Control oocytes remained untreated, whilst the remainder were exposed to three different combinations of vitrification solutions [dimethyl sulfoxide (DMSO) + ethylene glycol (EG), EG only, or propanediol (PROH) + EG] and either vitrified or left unfrozen (exposed groups). Oocytes in the control and vitrified groups underwent IVF and developmental competence was assessed to the blastocyst stage. In lambs, survival rate in vitrified oocytes was significantly lower than for oocytes in the exposed groups (P <0.05). Blastocyst development was low in vitrified oocytes compared with controls (<6% vs 38.9%, P <0.01). Parthenogenetic activation was more prevalent in vitrified lamb oocytes compared with controls (P <0.05). No evidence of zona pellucida hardening or cortical granule exocytosis could account for reduced fertilization rates in vitrified lamb oocytes. Mouse oocytes demonstrated a completely different response to lamb oocytes, with survival and parthenogenetic activation rates unaffected by the vitrification process. Treatment of mouse oocytes with DMSO + EG yielded significantly higher survival and cleavage rates than treatment with PROH + EG (87.8% and 51.7% vs 32.7% and 16.7% respectively, P <0.01), however cleavage rate for vitrified oocytes remained lower than for the controls (51.7% vs 91.7%, P <0.01) as did mean blastocyst cell number (33 ± 3.1 vs 42 ± 1.5, P <0.05). From this study, it is clear that lamb and mouse show different tolerances to cryoprotectants commonly used in vitrification procedures, and careful selection and testing of species-compatible cryoprotectants is required when vitrifying oocytes to optimize survival and embryo development.

Recent advances of oocyte/embryo vitrification in mammals from rodents and large animals

“…If vitrification becomes mainstream in cattle in the future, it can potentially be applied to other farm animals. As the tolerances to CPAs vary across species (Sudiman et al, 2019), it is challenging to apply effective methods in one animal to others. Specifically, the collapse of large blastocoels in horses (Benammar et al, 2021) and removal of lipid droplets in pigs (Nagashima et al, 1994(Nagashima et al, , 1995 necessitates the optimization of methods by taking into account the unique properties of each animal.…”

Recent advances of oocyte/embryo vitrification in mammals from rodents and large animals

“…If vitrification becomes mainstream in cattle in the future, it can potentially be applied to other farm animals. As the tolerances to CPAs vary across species (Sudiman et al, 2019), it is challenging to apply effective methods in one animal to others. Specifically, the collapse of large blastocoels in horses (Benammar et al, 2021) and removal of lipid droplets in pigs (Nagashima et al, 1994(Nagashima et al, , 1995 necessitates the optimization of methods by taking into account the unique properties of each animal.…”

Evaluation of chromosome organization and microtubule arrangement in goat (capra aegragrus) oocytes after vitrification, in vitro maturation and fertilization, and early embryo development

“…The vitrification method has different levels of success among oocytes of different species. In fact, the oocytes show different tolerances to the cryoprotectants used in the vitrification procedures (Sudiman et al, 2019). With regard to oocytes, the most reported alterations during cryopreservation are interruptions of the tubulin network, meiotic spindle disorganization, abnormal chromosomal distributions and a reduced in vitro fertilization rate (Tamura et al, 2013, Lei et al, 2014, Serra et al, 2020, probably caused by the increase in aneuploidy (Buderatska et al, 2020, Dviri et al, 2021, genomic alterations that entail structural (Gao et al, 2018, Wasielak-Politowska et al, 2022 and biochemical changes (Ren et al, 2019, Tsuiko et al, 2019 that compromise cell viability even to the point of cell death (Balboula et al, 2020, Vining et al, 2021.…”

Recent advances of oocyte/embryo vitrification in mammals from rodents and large animals

“…If vitrification becomes mainstream in cattle in the future, it can potentially be applied to other farm animals. As the tolerances to CPAs vary across species (Sudiman et al, 2019), it is challenging to apply effective methods in one animal to others. Specifically, the collapse of large blastocoels in horses (Benammar et al, 2021) and removal of lipid droplets in pigs (Nagashima et al, 1994(Nagashima et al, , 1995 necessitates the optimization of methods by taking into account the unique properties of each animal.…”

Evaluation of chromosome organization and microtubule arrangement in goat (capra aegragrus) oocytes after vitrification, in vitro maturation and fertilization, and early embryo development

“…The vitrification method has different levels of success among oocytes of different species. In fact, the oocytes show different tolerances to the cryoprotectants used in the vitrification procedures (Sudiman et al, 2019). With regard to oocytes, the most reported alterations during cryopreservation are interruptions of the tubulin network, meiotic spindle disorganization, abnormal chromosomal distributions and a reduced in vitro fertilization rate (Tamura et al, 2013, Lei et al, 2014, Serra et al, 2020, probably caused by the increase in aneuploidy (Buderatska et al, 2020, Dviri et al, 2021, genomic alterations that entail structural (Gao et al, 2018, Wasielak-Politowska et al, 2022 and biochemical changes (Ren et al, 2019, Tsuiko et al, 2019 that compromise cell viability even to the point of cell death (Balboula et al, 2020, Vining et al, 2021.…”

Recent advances of oocyte/embryo vitrification in mammals from rodents and large animals

“…If vitrification becomes mainstream in cattle in the future, it can potentially be applied to other farm animals. As the tolerances to CPAs vary across species (Sudiman et al, 2019), it is challenging to apply effective methods in one animal to others. Specifically, the collapse of large blastocoels in horses (Benammar et al, 2021) and removal of lipid droplets in pigs (Nagashima et al, 1994(Nagashima et al, , 1995 necessitates the optimization of methods by taking into account the unique properties of each animal.…”

Evaluation of chromosome organization and microtubule arrangement in goat (capra aegragrus) oocytes after vitrification, in vitro maturation and fertilization, and early embryo development

“…The vitrification method has different levels of success among oocytes of different species. In fact, the oocytes show different tolerances to the cryoprotectants used in the vitrification procedures (Sudiman et al, 2019). With regard to oocytes, the most reported alterations during cryopreservation are interruptions of the tubulin network, meiotic spindle disorganization, abnormal chromosomal distributions and a reduced in vitro fertilization rate (Tamura et al, 2013, Lei et al, 2014, Serra et al, 2020, probably caused by the increase in aneuploidy (Buderatska et al, 2020, Dviri et al, 2021, genomic alterations that entail structural (Gao et al, 2018, Wasielak-Politowska et al, 2022 and biochemical changes (Ren et al, 2019, Tsuiko et al, 2019 that compromise cell viability even to the point of cell death (Balboula et al, 2020, Vining et al, 2021.…”