Impact of Presence or Absence of Trehalose during Vitrification on Viability and Development of Vitrified/Warmed Immature Dromedary Camel Oocytes

Yaqout, Karim A; Monir, Ahmed; Badr, M. A.; Elwishy, A.B.; Moawad, Adel R.; El-Shalofy, Amr S.

doi:10.4236/ojas.2022.123026

Cited by 2 publications

(2 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One of the main reasons contributing to this scenario is the confined availability of oocytes due to the shortage of abattoirs slaughtering female camels as well as the lack of a standardized protocol for IVM and IVF of dromedary camel oocytes. Cryopreservation of oocytes would be an alternative way to circumvent the limited availability of oocytes, allowing for improvements in IVEP in this species (Abri et al, 2019;Fathi et al, 2018Fathi et al, , 2021Moawad et al, 2020;Yaqout et al, 2022). A previous study reported that dromedary camel oocytes vitrified at the germinal vesicle stage could be matured, fertilized and subsequently cultured in vitro up to blastocyst stages; however, the cleavage (22.5%-27.9%), morula (13.2%-14.5%) and blastocyst rates (6.4%-8.5%) rates were low compared with those reported to other domestic species (Fathi et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Influences of glycine supplementation during vitrification on the developmental potential of vitrified/warmed immature dromedary camel oocytes

Yaqout

Bard

Elwishy

et al. 2023

Reprod Domestic Animals

Self Cite

View full text Add to dashboard Cite

Oocytes experience detrimental osmotic stress during vitrification and warming procedures because of the osmolality imbalance between the vitrification‐warming fluids and the intracellular environment. Cellular osmotic homeostasis can be preserved by glycine, a powerful osmolyte with antioxidant properties. We aimed to examine the influences of supplementing glycine to the vitrification solutions (VS) on the developmental potential of vitrified/warmed immature dromedary camel oocytes following IVM/IVF and in vitro embryo culture (IVC). Cumulus oocyte complexes (COCs) were collected from dromedary camel ovaries and randomly allocated into two groups namely control (oocytes subjected directly to IVM) and vitrified (COCs were vitrified into VS supplemented with 0.0, 0.5, 1.0 or 2.0 mM glycine). For vitrification, COCs were equilibrated for 3 min in 12.5% ethylene glycol; EG plus 12.5% dimethyl sulfoxide; DMS and then they were vitrified for 60 s in VS composed of 25% EG + 25% DMSO using solid surface vitrification (SSV). Warming of vitrified oocytes was conducted in decreasing concentrations of trehalose solution. Following vitrification and warming, the morphologically viable oocytes were subjected to IVM for 36 h. Matured oocytes were then fertilized in vitro by epididymal spermatozoa and cultured for seven days. The results showed that the percentage of viable oocytes assessed by trypan blue stain was significantly higher (p ≤ .05) in the 1.0 mM glycine‐supplemented group than 0.0‐ and 2.0‐mM glycine‐supplemented ones (90.0 % vs. 80.0% and 76.6%, respectively). However, no significant difference was observed between 0.5 mM glycine and other vitrified groups. Nuclear maturation rates, cleavage (48‐h post‐insemination; pi) and blastocyst rate (7‐days pi) were significantly lower in vitrified groups than control ones (p ≤ .05). Among vitrified groups, these parameters were the highest in the 1.0 mM glycine‐supplemented group. Taken together, supplementation of vitrification solutions with 1.0 mM glycine could enhance the developmental potential of vitrified/warmed immature dromedary camel oocytes.

show abstract

Section: Introductionmentioning

confidence: 99%

Influences of glycine supplementation during vitrification on the developmental potential of vitrified/warmed immature dromedary camel oocytes

Yaqout

Bard

Elwishy

et al. 2023

Reprod Domestic Animals

Self Cite

View full text Add to dashboard Cite

show abstract

“…Cryoprotectants and vitrification have been extensively used for humans (Fabbri, 2006) as well as other species, such as porcine (Zhou and Li, 2009), bovine (Hwang and Hochi, 2014) and goat (Purohit et al, 2012). Some recent studies on the verification of immature camel oocytes discovered that vitrification caused mechanical damage and reduces the potential for oocyte development (Moawad et al, 2019;Yaqout et al, 2022). Moreover, vitrification has adverse effects on mitochondrial functions, gene transcript, and camel oocyte development (Saadeldin et al, 2020;Moulavi et al, 2021).…”

mentioning

confidence: 99%

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Kandil¹,

Aboelwafa²,

Ismail³

et al. 2022

WVJ

View full text Add to dashboard Cite

In vitro embryo generation, cryopreservation, and embryo transfer are examples of assisted reproductive technologies that can be used to improve camel genetic performance and fertility. The aim of this study was to investigate the impact of ascorbic acid supplementation to in vitro maturation media on the maturation rate, morphology, and gene expression of fresh and vitrified in vitro matured dromedary camel oocytes. In the current study, 810 oocytes of excellent and good quality were in vitro matured in maturation medium (TCM-199 + 10 ug/ml follicle stimulated hormone + 10% fetal calf serum + 100 IU/ml Pregnant mare serum + 50 μg/ml gentamycin) without any additives to act as a control group (C) and with 50 μg/ml ascorbic acid group (AA) and incubation in a CO2 incubator (38.5 ̊C, 5% CO2, 20% O2 and 95% humidity) for 40 hours. In vitro matured dromedary camel oocytes with the first polar body (n = 210) in C group and AA group (n = 250) were placed in basic medium (BM) and then placed in vitrification solution1 (VS1) for one minute, followed by the transfer of oocytes to VS2 (double concentration of VS1, containing 20% Ethyl Glycol (EG) and +20% Dimethyl sulfoxide) for 30 seconds. Oocytes were then loaded into sterile 0.25 ml straws and stored in liquid nitrogen for 7-10 days. The normal fresh and vitrified /thawed in vitro matured dromedary camel oocytes were kept in RNA later at a -80°C freezer for gene expression analysis. The maturation rate of dromedary camel oocytes in the in vitro matured AA group was significantly higher than that of the C group. The percentage of normally recovered vitrified/thawed oocytes was higher in the in vitro matured with ascorbic acid (VAA) than in the control (VC) group. The expression pattern of the SOD1 gene and GDF9 gene was upregulated in fresh AA and VAA groups than in the fresh C and VC groups. The profile of the SOD1 gene was more abundant in the vitrified/thawed oocytes VAA group than in the VC group. All vitrified/thawed groups, whether control or ascorbic acid supplemented, had lower levels of SOD1, GDF9, and BMP15 expression, compared to the fresh groups. In conclusion, the supplementation of the maturation medium with ascorbic acid has an increased maturation rate, and normal morphology of vitrified/ thawed oocytes which was linked with upregulation of SOD1, GDF9 genes expression.

show abstract

Impact of Presence or Absence of Trehalose during Vitrification on Viability and Development of Vitrified/Warmed Immature Dromedary Camel Oocytes

Cited by 2 publications

References 38 publications

Influences of glycine supplementation during vitrification on the developmental potential of vitrified/warmed immature dromedary camel oocytes

Influences of glycine supplementation during vitrification on the developmental potential of vitrified/warmed immature dromedary camel oocytes

Effects of Ascorbic Acid on Maturation Rate, Morphology, and Gene Expression of Vitrified In Vitro Matured Dromedary Camel Oocytes

Contact Info

Product

Resources

About