The Role of Forskolin as a Lipolytic Stimulator during In Vitro Oocyte Maturation and the in Vitro Embryo Production of Livestock

2021 ◽  
Author(s):  
Sayed Haidar Abbas Raza ◽  
Ayman H. Abd El‐Aziz ◽  
Sameh A. Abdelnour ◽  
Ahmed A. Easa ◽  
Mahmoud Alagawany ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
In Vitro Oocyte Maturation

PSX-A-23 Late-Breaking: Supplementation of Nerve Growth Factor (β-NGF) in the maturation medium and efficiency of in vitro embryo production in cattle

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 365-365
Author(s):  
Lucas Gonçalves ◽  
Muller C Martins ◽  
Natalia Arle ◽  
Rafaela T Torres ◽  
Luisa Migilo ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Oocyte Maturation ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Nerve Growth ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Abstract The aim of this study was to evaluate the supplementation of Nerve Growth Factor (β-NGF) in the maturation medium in in vitro embryo production routines. Antral follicles were aspirated from ovaries of cows obtained from slaughterhouses and then oocytes were selected for quality (grades I and II) for in vitro maturation and subjected to 4 successive in vitro embryo production routines (IVEP). Supplementation of 100 ng of β-NGF was performed in the oocyte maturation medium 22 hours before in vitro fertilization. 48 hours after fertilization of the oocytes, an analysis was made of their cleavage rate by counting blastomeres with the aid of a stereoscopic microscope (cleavage rate = number of embryos / number of initial oocytes). Seven days after fertilization, the blastocyst rate was determined through the relation to the number of oocytes that started cleavage and reached this stage of development (blastocyst rate = number of blastocyst / number of oocytes that started cleavage). To verify the existence of a difference between the supplemented and the non-supplemented groups, the paired T test was applied, using the Excel / Action software (Microsoft). In vitro embryo production routines supplemented with β-NGF in the maturation medium had, on average, a higher cleavage rate (P = 0.0072) and a higher blastocyst rate (P = 0.0033) compared to non-supplemented routines with β-NGF. In this study was demonstrated that Nerve Growth Factor supplementation in the maturation medium improves the efficiency of in vitro embryo production in cattle, and this protein has a probable action in the oocyte maturation process.

scholarly journals Exposure to follicular fluid during oocyte maturation and oviductal fluid during post-maturation does not improve in vitro embryo production in the horse

Zygote ◽  
2017 ◽  
Vol 25 (5) ◽  
pp. 612-630 ◽  
Author(s):  
Cécile Douet ◽  
Olivia Parodi ◽  
Nicola Antonio Martino ◽  
Giovanni Michele Lacalandra ◽  
Michele Nicassio ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Culture Conditions ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Oviductal Fluid

SummaryMost wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) technique for equine embryo production is available. The development of culture conditions able to mimic the maturation of the oocyte in preovulatory follicular fluid (pFF) and the post-maturation in oviductal fluid (OF) may improve embryo production in vitro. Our aim was to analyse the effect of in vitro maturation in pFF and incubation in OF on in vitro maturation of equine oocytes, fertilization using conventional IVF or ICSI, and embryo development after culture in synthetic oviductal fluid (SOF) or DMEM-F12. Oocytes collected from slaughtered mares or by ovum pick up were matured in vitro in pFF or semi-synthetic maturation medium (MM). The in vitro maturation, fertilization and development rates were not statistically different between pFF and MM. After in vitro maturation, oocytes were incubated with or without OF. Post-maturation in OF did not significantly improve the fertilization and development rates. Thus, in our study, exposure to physiological fluids for oocyte maturation and post-maturation does not improve in vitro embryo production in the horse.

State-of-the-art production, conservation and transfer of in-vitro-produced embryos in small ruminants

2004 ◽  
Vol 16 (4) ◽  
pp. 437 ◽  
Author(s):  
Yves Cognié ◽  
Nati Poulin ◽  
Yann Locatelli ◽  
Pascal Mermillod
Keyword(s):  
Oocyte Maturation ◽  
Small Ruminants ◽  
Reproductive Technologies ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Embryo Production ◽  
Oocyte Competence ◽  
Epidermal Growth ◽  
In Vitro Embryo Production

Today, although not efficient enough to replace multiple ovulation and embryo transfer, in vitro embryo production for small ruminants is a platform for new reproductive technologies, such as embryo sexing, transgenesis and cloning. The in vitro embryo-production system developed for sheep and goats is more efficient now than 15 years ago, but could still be improved. Laparoscopic collection of oocytes in live animals treated with gonadotrophin indicates a promising future for the application of this technology to genetic improvement programmes. Oocyte maturation in defined medium with epidermal growth factor and cysteamine appears as efficient as oocyte maturation in follicular fluid-supplemented medium and allows future study of the effect of other factors involved in the cytoplasmic maturation of oocytes from these species. Further efforts have to be made to standardise the semen-capacitating process and to improve the quality and freezability of in-vitro-produced (IVP) embryos. The optimisation of IVP procedures for deer species has required the study of the seasonal variation of oocyte competence and the development of a specific methodology to allow the culture of embryos up to the blastocyst stage.

74 Follicular fluid anti-Müllerian hormone concentration predicts juvenile ovine in vitro embryo development

2019 ◽  
Vol 31 (1) ◽  
pp. 162
Author(s):  
J. E. Seccafien ◽  
J. M. Kelly ◽  
H. McGrice ◽  
D. O. Kleemann ◽  
K. L. Kind ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Individual Variability ◽  
Developmental Competence ◽  
Response To Treatment ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Oocyte Developmental Competence ◽  
In Vitro Embryo Production

Currently, the commercial viability of assisted reproductive embryo technologies within the Australian livestock industry is restricted by individual variability in response to treatment protocols as well as oocyte developmental competence. The majority of losses come from embryo wastage, resulting from poor developmental competence during in vitro embryo production. Follicular fluid is readily available when oocytes are collected for in vitro embryo production from juvenile or mature ewes, making it an appropriate target for analysis of phenotypic markers of oocyte developmental competence. Plasma anti-Müllerian hormone (AMH) is correlated with pregnancy losses, oocyte recovery, and blastocyst development in sheep and cattle and is an indicator for donors that respond best to gonadotrophin stimulation protocols in sheep, cattle, and goats. The aim of the current work was to determine the relationship between follicular fluid AMH and in vitro embryo production outcomes in sheep. Briefly, pairs of ovaries from 38 abattoir-derived lambs were collected individually and transferred to the laboratory. Ovaries were aspirated for in vitro embryo production following previously described methods (Walker et al. 1996 Biol. Reprod. 55, 703-708) and follicles counted. Aspirated oocytes from each of the 38 individual lamb’s pair of ovaries were pooled [n=4.11±0.53 cumulus-oocyte complexes (COC) matured/lamb; total COC matured=156], and remained as such during maturation, fertilisation, and culture. The remaining follicular fluid was centrifuged for 10min at 3000 rpm to remove excess cells and frozen at −20°C. The AMH was measured in follicular fluid by a human AMH Gen II ELISA kit validated for ovine samples (A79766, Beckman Coulter, Brea, CA, USA). Correlations between follicular fluid AMH levels and oocyte maturation and blastocyst development were determined using simple linear regression. Animals were divided into groups based on AMH levels [low (0.5-10.8ng mL−1), medium (10.81-17.89ng mL−1), or high (17.9-19.25ng mL−1)], with an unbalanced ANOVA used to determine group effects on oocyte maturation and blastocyst development (GenStat 18th edition, VSN International, Hemel Hempstead, UK). Follicular fluid AMH was positively correlated (P<0.05) with the number of follicles greater than 2mm (r2=0.120) and the proportion of COC cleaved from recovered oocytes (r2=0.134). The number of COC matured per lamb was greater for those with high and medium versus low AMH (5.6±0.97 and 4.4±0.72 versus 2.1±0.97 COC/lamb). Animals with high AMH produced more blastocysts than those with medium or low AMH, when expressed as a proportion of COC recovered (P<0.002) or cleaved (P<0.009) oocytes. High AMH was also correlated with a greater number of expanded blastocysts produced from cleaved oocytes (P<0.042). The current data support previous evidence that AMH levels positively correlate to higher antral follicle counts. The correlation between AMH and components of oocyte developmental competence suggests intrafollicular AMH may indicate the best oocytes to use for an in vitro embryo production system.

scholarly journals In vitro embryo production in small ruminants

2010 ◽  
Vol 39 (suppl spe) ◽  
pp. 409-413 ◽  
Author(s):  
Vicente José de Figueirêdo Freitas ◽  
Luciana Magalhães Melo
Keyword(s):  
Molecular Biology ◽  
Oocyte Maturation ◽  
Genetic Improvement ◽  
In Vitro Maturation ◽  
Small Ruminants ◽  
Northeastern Brazil ◽  
Improvement Program ◽  
Embryo Production ◽  
In Vitro Embryo Production

This paper reviews the technical bases of in vitro embryo production in small ruminants with special attention to the results obtained by our group in Northeastern Brazil. The laparoscopic oocyte recovery in hormonally treated live animals indicates a promising future for the application of this technique to genetic improvement program. New molecular biology tools should provide information to improve the efficiency of in vitro maturation. Furthers efforts have to be made to improve the oocyte maturation and to standardize the semen-capacitating process.

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