In Vitro Maturation, Fertilization and Development of Prepubertal and Mature Buffalo Oocytes

Kadoom, A. K.; Abdel-Khalek, A. E.; Shamiah, Sh. M.; El–Sharawy, Mohamed; El-Razek, Ibrahim M. Abd

doi:10.21608/ejap.2014.93652

Cited by 3 publications

(1 citation statement)

References 24 publications

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“…Selected good COCs were washed four times in fresh warm aspiration medium and subjected to the final wash using an IVM medium. In 35-mm Petri dishes, groups of 5–15 COCs were cultured in 50–100 μL droplets of the filtrated IVM media (pH 7.4) covered with sterilized mineral oil for 24 h at 5% CO 2 , 38.5°C, and 90–95% relative humidity ( 25 ).…”

Section: Methodsmentioning

confidence: 99%

Impact of epidermal growth factor and/or β-mercaptoethanol supplementations on the in vitro produced buffaloes' embryos

et al. 2023

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The present study investigated the effects of epidermal growth factors (EGF) and/or β-Mercaptoethanol (βME) supplementations to oocyte maturation, fertilization, and culture media on the buffalo in vitro embryo production. The ovaries were collected and transferred within 2 h to the laboratory. The cumulus oocytes complexes were aspirated from 3 to 8 mm diameter follicles. Firstly, EGF; 0, 10, 20, or 50 ng/mL or βME; 0, 25, 50, 100, or 200 μM were supplemented to the in vitro maturation (TCM-199), fertilization (IVF-TALP), or culture (IVC: SOF) media. Our results revealed that supplementing EGF (20 ng/mL) to the TCM-199, IVF-TALP, or SOF media could efficiently improve the growth rates and development of buffalos' embryos, while EGF (50 ng/mL) could stimulate the embryo production only after treatment of the IVF-TALP /or SOF media, but not the IVM medium. However, βME was less efficient than EGF; it stimulated the growth rates of buffalo embryos when supplemented with the maturation and fertilization (IVF-TALP) media in a 50 μM concentration. Secondly, combined EGF (20 ng/mL) and βME (50 μM) were supplemented to the maturation media as effective concentration. The combined treatment of EGF (20 ng/mL) and βME (50 μM) showed no significant enhancing effect on the buffalo embryos compared to each alone. For future perspectives, further study is required to examine the effects of combined EGF and βME on the maturation and fertilization of buffalo oocytes at different categories of age and seasonal localities.

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Section: Methodsmentioning

confidence: 99%

Impact of epidermal growth factor and/or β-mercaptoethanol supplementations on the in vitro produced buffaloes' embryos

et al. 2023

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Cleavage and in vitro cultivation rates monitoring in culture media supplemented with energy sources, non-essential amino acids, and antioxidants in the buffalo embryos

Abu El-Naga,

Ali,

Ali

et al. 2024

BMC Vet Res

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The study was designed to monitor the cleavage rate (CR) and in-vitro cultivation rate (IVC) after addition of energy sources, non-essential amino acids, and antioxidants to the Synthetic oviductal fluid (SOF) and FertiCult. After in-vitro maturation and in-vitro fertilization, presumptive zygotes were cultured in one of two culture media: FertiCult media and SOF medium, supplemented with pyruvate, glucose, and sodium lactate as energy sources, as well as 10, 20, 250, 500, and 750 mg non-essential amino acids, and antioxidants. All stages of cleavage rate (CR), and in-vitro cultivation rate (IVC) of embryonic development including morula stage (MOR) and blastocyst (BLAS) have been assessed. The findings revealed that there were no significant differences in the CR between the control and other treated groups with sources of energy when added to SOF media (P > 0.05), while there were significant differences (P < 0.05) in the IVC of embryonic development between groups (The percentages of MOR stage in the control, pyruvate, glucose and mixture of source of energy (MIX) were at 50%, 62.5%, 60%, and 63.6%, respectively). The highest percentage of the BLAS was recorded after SOF supplementation with glucose (40%). Similarly, there were no significant differences (P > 0.05) in the CR between control and FertiCult supplemented with sources of energy, while the IVC stages increased significantly (P < 0.05) in the FertiCult media supplemented with glucose, pyruvate, sodium lactate, and MIX. The percentages of the MOR stage in the control, pyruvate, glucose and mix media were at 50%, 55.6%, 55.6%, 54.5%, 57.1% respectively. The lowest percentage of the BLAS was recorded after FertiCult supplementation with pyruvate (11.1%). Replenishing the SOF maturation media with 20 mg of non-essential amino acids significantly (P < 0.05) enhanced the MOR stage (100%). There was also an improvement in the development of BLAS stage, where it reached 31.2% and 47.4% in the SOF maturation media supplemented with 10, and 750 mg non-essential amino acids, respectively. There were no significant differences (P > 0.05) in neither CR nor IVC between control and FertiCult supplemented with antioxidants. There were significant differences (P < 0.05) in the MOR stages (control, 42.9% & treated, 57.9%) and BLAS stages (control, 21.4% & treated, 42.1%) in antioxidant supplemented SOF maturation media compared to control. In conclusion, supplementation of SOF cultivation medium with energy sources, 20 mg of non-essential amino acids and antioxidant addition may improve the cleavage rate (CR) and in vitro cultivation rate (IVC) of buffalos’ embryonic development.

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In vitro maturation of collared peccary (Pecari tajacu) oocytes after different incubation times

et al. 2018

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Oocyte in vitro maturation (IVM) is the first step of the in vitro reproductive technologies that enables mature oocytes to be generated ex vivo and after used for embryo production. In this sense, the establishment of culture environment, as oocyte incubation time, is essential for the success of the IVM. Therefore, the study was carried out to investigate the relationship between the meiotic potential and the IVM times of collared peccary oocytes, wild mammals of great commercial and ecological interest. Thus, ovaries were collected of females derived from captivity and transported to the laboratory within 1 hour of slaughtering. The oocytes derived from follicles (3-6mm in diameter) were recovered by aspirated and sliced. Good quality oocytes (evenly granulated cytoplasm with a least one layer of surrounding cumulus cells) were selected and subjected to culture in TCM 199 supplemented with 10µg/mL FSH, 10% FBS and 100µM cysteamine at 38.5°C, 5% CO2 and maximum humidity for 24 or 48 hours. After the incubation period, the nuclear status, the presence of first polar body and the expansion of cumulus cells of oocytes were assessed. The data obtained were analyzed by Fisher exact test (P<0.05). A total of four sessions (2-3 females per session) were performed, resulting in eighteen aspirated and sliced ovaries with normal morphological characteristics. An oocyte recovery rate of about 83.1% (59/71) was obtained with 3.3 oocytes/ovary and 2.3 viable oocytes/ovary. After different incubation times, differences (P<0.05) were observed in 24 and 48 hours for expansion of the cumulus cells (38.1% vs. 100%), presence of first polar body (52.4% vs. 90.5%) and nuclear status in second metaphase (19.0% vs. 76.2%), respectively. In conclusion, 48 hours is suitable time for the in vitro maturation of oocytes derived from collared peccaries when compared to the time of 24 hours, according to the meiotic potential observed. Additional studies should be conducted to improve the quality of the oocyte culture environment, as medium composition, aiming to obtain viable mature oocytes for other in vitro biotechnologies.

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In Vitro Maturation, Fertilization and Development of Prepubertal and Mature Buffalo Oocytes

Cited by 3 publications

References 24 publications

Impact of epidermal growth factor and/or β-mercaptoethanol supplementations on the in vitro produced buffaloes' embryos

Impact of epidermal growth factor and/or β-mercaptoethanol supplementations on the in vitro produced buffaloes' embryos

Cleavage and in vitro cultivation rates monitoring in culture media supplemented with energy sources, non-essential amino acids, and antioxidants in the buffalo embryos

In vitro maturation of collared peccary (Pecari tajacu) oocytes after different incubation times

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