Morphometric Assessment of the Bovine Ovary for in vitro Matured Oocyte Quality to Determine Developmental Competence

Ammari, Aiman A.; Amran, Ramzi A.; Ghadi, Muath G. Al; Alhimaidi, Ahmad R.

doi:10.18805/ijar.bf-1471

Cited by 5 publications

(10 citation statements)

References 28 publications

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“…Inc., Smithfield, UT, USA)). After maturing in the lab for 24 h, cumulus–oocyte complex (1832) showed uniform cytoplasm and more than three layers of cumulus cells [ 50 ].…”

Section: Methodsmentioning

confidence: 99%

Interspecific Nuclear Transfer Blastocysts Reconstructed from Arabian Oryx Somatic Cells and Domestic Cow Ooplasm

Ammari

Al-Ghadi

Amran

et al. 2022

Veterinary Sciences

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Cloning, commonly referred to as somatic cell nuclear transfer (SCNT), is the technique of enucleating an oocyte and injecting a somatic cell into it. This study was carried out with interspecific SCNT technology to clone the Arabian Oryx utilizing the oryx’s fibroblast cells and transfer it to the enucleated oocytes of a domestic cow. The recipient oocytes were extracted from the cows that had been butchered. Oryx somatic nuclei were introduced into cow oocytes to produce embryonic cells. The study was conducted on three groups, Oryx interspecific somatic cell nuclear transfer into enucleated oocytes of domestic cows, cow SCNT “the same bovine family species”, used as a control group, and in vitro fertilized (IVF) cows to verify all media used in this work. The rates of different embryo developmental stages varied slightly (from 1- cell to morula stage). Additionally, the oryx interspecies Somatic cell nuclear transfer blastocyst developmental rate (9.23%) was comparable to that of cow SCNT (8.33%). While the blastula stage rate of the (IVF) cow embryos exhibited a higher cleavage rate (42%) in the embryo development stage. The results of this study enhanced domestic cow oocytes’ ability to support interspecific SCNT cloned oryx, and generate a viable embryo that can advance to the blastula stage.

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“…Inc., Smithfield, UT, USA)). After maturing in the lab for 24 h, cumulus–oocyte complex (1832) showed uniform cytoplasm and more than three layers of cumulus cells [ 50 ].…”

Section: Methodsmentioning

confidence: 99%

Interspecific Nuclear Transfer Blastocysts Reconstructed from Arabian Oryx Somatic Cells and Domestic Cow Ooplasm

Ammari

Al-Ghadi

Amran

et al. 2022

Veterinary Sciences

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“…Next the cumulus-oocyte complexes underwent three washings in 100 μl of TCM-199 maturation medium (Earle's salts) (Sigma M4530; Merck KGaA, Darmstadt, Germany) with 0.02 IU/ml follicle-stimulating hormone (FSH) (Sigma F8174), 0.023 IU/ml luteinizing hormone (LH) (Sigma L5269), 1 μg/ml Estradiol (E2) (Sigma E2758), 10% FBS, 50 μg/ml gentamicin sulfate and 0.5 mM Na-Pyruvate. After being separated into ten groups, the cumulus-oocyte complexes were placed in 50 ml maturation drops in 60 mm Petri dishes, coated with mineral oil and incubated at 39  C in a humidified atmosphere of 5% CO 2 for 24 hours (Ammari et al, 2022a;Ammari et al, 2022b).…”

Section: Collection Of Oocytes and In Vitro Maturationmentioning

confidence: 99%

“…The oocytes were four times washed in the preceding capacitation medium (BO-IVF) before transferring them to the fertilization drops. The fertilization medium with 10 oocytes per 50 μl drop was inseminated with 2×10 6 sperm and incubated at 39  C in a humidified environment with 5% CO 2 for 24 hours (Amran et al, 2023).…”

Section: In Vitro Fertilization Of Oocytesmentioning

confidence: 99%

“…The zygotes were cultured in a 35 mm Petri plate (BD Falcon, 351008) with 50 μl drops of the SOF medium and 25 embryos in each drop, followed by the addition of mineral oil (Sigma M 5310). Embryos were incubated for 7 to 8 days at 39  C with 5% CO 2 , 90% N2 and 5% O 2 in a highly humid atmosphere (Ammari et al, 2022a;Ammari et al, 2022b;Amran et al, 2023).…”

Section: In Vitro Culture Of Embryosmentioning

confidence: 99%

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Impact of Ionomycin on the Gene Expression of In vitro Fertilized Bovine Embryos

Amran,

Alhimaidi,

Ammari

et al. 2024

IJAR

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Background: The embryo genomic material used in this study came from an equal contribution of maternal and paternal genes. Gene expression during IVEP embryo development was controlled by several factors: the genetic makeup of the embryo, the epigenetic contributions and the culture materials. The aims of this research were to study the impact of ionomycin treatment on the sperm capacitation time and its impacts on the gene expression of some sensitive genes (Bcl-2, Bax, HSPB1, IGF1 and GDF9) during IVEP. Methods: The semen was placed in a sterile 15ml tube containing 5ml of capacitation medium and then split into three different experimental groups: the first group was the control; the second was the low-treated group, which contained sperm in a capacitation medium with 25 nM of ionomycin; the third group included 50 nM ionomycin. Result: The gene expression of BAX during early cleavage did not show any differences. However, in the late cleavage stage, the control group presented higher gene expression (P less than 0.05) than that in the two treated groups. The Bcl-2 genes of the control group showed higher gene expression (P less than 0.05) in both the early and late cleavage stages compared to the low-treated group. Gene expression of the HSP-1 gene was observed during early embryo development in the treated group (25 nM ionomycin). In conclusion, concentration 25 nM ionomycin showed higher gene expression in the early stages of embryo development also the best-quality embryos and gene expression results were found by studying gene expression. The transition from the maternal and paternal genome to the embryonic genome and gene expression during embryo development is controlled by several factors.

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“…Then, groups of ten cumulus-oocyte complexes were transferred to 60 mm Petri dishes in 50 μl maturation drops which were covered with mineral oil and incubated for 24 hours in a humidified atmosphere of 5% CO 2 at 39C. (Ammari et al, 2022a).…”

Section: Oocyte Collection and In Vitro Maturation (Ivm)mentioning

confidence: 99%

The Role of Ionomycin in the Improvement of in vitro Fertilization and Increasing the Rate of Bovine Embryo Development

Amran¹,

Aleissa²,

Ghadi³

et al. 2023

IJAR

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Background: The utilization of in vitro fertilization (IVF) and in vitro embryo production (IVEP) has increased significantly as an effective tool of assisted reproductive technology. Sperm capacitation is a process of membrane maturation allowing the acrosome reaction to take place. Ionomycin is a calcium ionophore that induces the acrosome reaction by increasing calcium cycling across phospholipid membranes. The aim of this study was to enhance the cleavage rate and development of bovine embryos. Methods: Bovine ovaries were collected from the central slaughter house. Oocytes were obtained from follicles and were let to mature for 24 h. Frozen bull semen thawed in 37°C water and transferred to a discontinuous percoll gradient (Sigma GE17-0891-01). The spermatozoon pellet was separated into four groups after resuspension in bovine in vitro fertilization media (BO-IVF): control without additives; with DMSO and with ionomycin (25 and 50 nM) and kept in an incubator for three hours. The oocytes matured under commonly used media (TCM-199) for 24 h. The mature oocytes were then fertilized for 18-24h. The embryos were cultured in SOF medium until the seventh day to study cleavage rate and blastocyst rate. Result: The results indicated that 50 nM ionomycin supplementation considerably reduced cleavage while DMSO and 25 nM ionomycin supplementation had no effect on cleavage percentage. Moreover, when compared to the other groups, ionomycin at 25 nM significantly increased blastocyst development. In conclusion, the highest rate of blastocyst development was observed in the capacitation medium supplemented with 25 nM ionomycin, apparently due to the importance of calcium and metabolism in spermatozoa for fertilization and embryo development.

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Morphometric Assessment of the Bovine Ovary for in vitro Matured Oocyte Quality to Determine Developmental Competence

Cited by 5 publications

References 28 publications

Interspecific Nuclear Transfer Blastocysts Reconstructed from Arabian Oryx Somatic Cells and Domestic Cow Ooplasm

Interspecific Nuclear Transfer Blastocysts Reconstructed from Arabian Oryx Somatic Cells and Domestic Cow Ooplasm

Impact of Ionomycin on the Gene Expression of In vitro Fertilized Bovine Embryos

The Role of Ionomycin in the Improvement of in vitro Fertilization and Increasing the Rate of Bovine Embryo Development

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