Factors affecting bovine embryo development in synthetic oviduct fluid following oocyte maturation and fertilization in vitro

Ovine embryos produced in synthetic oviduct fluid (SOF) medium or in coculture with granulosa cell monolayers supplemented with low (A; 120 μmol/l) and high (B; 190 μmol/l) ammonia-producing steer sera caused different degrees of fetal oversize (Carolan et al., 1998). The objective of the present study was to determine whether the effects on fetal growth induced by these sera were associated with alterations in early embryo development.A total of 911 bovine oocytes, used in 8 replicates to test the effect of three culture treatments on embryo development, were matured and fertilized in vitro (IVF= Day 0). Presumptive zygotes were allocated on Day 1 to culture in SOF supplemented with 10% v/v steer serum (SOF+A, n=308; SOF+B, n=302) or with amino acids plus 0.4% w/v crystalline BSA (SOFaaBSA, n=301). All cultures were in 20 μl droplets under oil (38.5°C; 5% CO2, 5% O2; 4 zygotes per drop) and droplets were renewed every 48 h. Cleavage rate was recorded on Day 3. On Days 7 and 8, blastocyst yields, grade 1 and 2 blastocysts, their cell numbers (by staining with Hoechst 33342) and their stage and diameter were determined.

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Effect of leptin supplementation during in vitro oocyte maturation and embryo culture on bovine embryo development and gene expression patterns

Theriogenology ◽

10.1016/j.theriogenology.2010.10.031 ◽

2011 ◽

Vol 75 (5) ◽

pp. 887-896 ◽

Cited By ~ 29

Author(s):

M. Arias-Alvarez ◽

P. Bermejo-Alvarez ◽

A. Gutierrez-Adan ◽

D. Rizos ◽

P.L. Lorenzo ◽

...

Keyword(s):

Gene Expression ◽

Embryo Development ◽

Oocyte Maturation ◽

Embryo Culture ◽

Expression Patterns ◽

Gene Expression Patterns ◽

Bovine Embryo ◽

In Vitro Oocyte Maturation

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Exogenous paraoxonase-1 during oocyte maturation improves bovine embryo development in vitro

Reproduction in Domestic Animals ◽

10.1111/rda.12730 ◽

2016 ◽

Vol 51 (5) ◽

pp. 827-830 ◽

Cited By ~ 8

Author(s):

JAA Rincón ◽

EM Madeira ◽

FT Campos ◽

B Mion ◽

JF Silva ◽

...

Keyword(s):

Embryo Development ◽

Oocyte Maturation ◽

Paraoxonase 1 ◽

Bovine Embryo ◽

Development In Vitro

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Effect of butafosfan supplementation during oocyte maturation on bovine embryo development

Zygote ◽

10.1017/s0967199419000327 ◽

2019 ◽

Vol 27 (05) ◽

pp. 321-328

Author(s):

Lucas Teixeira Hax ◽

Joao Alveiro Alvarado Rincón ◽

Augusto Schneider ◽

Lígia Margareth Cantarelli Pegoraro ◽

Letícia Franco Collares ◽

...

Keyword(s):

Gene Expression ◽

Embryo Development ◽

Oocyte Maturation ◽

Cumulus Cells ◽

Oocyte Quality ◽

Bovine Embryo ◽

Cleavage Rate ◽

Nuclear Maturation

SummaryAround 60–80% of oocytes maturated in vivo reached competence, while the proportion of maturation in vitro is rarely higher than 40%. In this sense, butafosfan has been used in vivo to improve metabolic condition of postpartum cows, and can represent an alternative to increase reproductive efficiency in cows. The aim of this study was to evaluate the addition of increasing doses of butafosfan during oocyte maturation in vitro on the initial embryo development in cattle. In total, 1400 cumulus–oocyte complexes (COCs) were distributed in four groups and maturated according to supplementation with increasing concentrations of butafosfan (0 mg/ml, 0.05 mg/ml, 0.1 mg/ml and 0.2 mg/ml). Then, 20 oocytes per group were collected to evaluate nuclear maturation and gene expression on cumulus cells and oocytes and the remaining oocytes were inseminated and cultured until day 7, when blastocysts were collected for gene expression analysis. A dose-dependent effect of butafosfan was observed, with decrease of cleavage rate and embryo development with higher doses. No difference between groups was observed in maturation rate and expression of genes related to oocyte quality. Our results suggest that butafosfan is prejudicial for oocytes, compromising cleavage and embryo development.

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Oocyte maturation under lipotoxic conditions induces carryover transcriptomic and functional alterations during post-hatching development of good-quality blastocysts: novel insights from a bovine embryo-transfer model

Human Reproduction ◽

10.1093/humrep/dez248 ◽

2020 ◽

Vol 35 (2) ◽

pp. 293-307

Author(s):

Karolien L J Desmet ◽

Waleed F A Marei ◽

Christophe Richard ◽

Katrien Sprangers ◽

Gerrit T S Beemster ◽

...

Keyword(s):

Gene Expression ◽

Rna Sequencing ◽

Embryo Transfer ◽

Embryo Development ◽

Oocyte Maturation ◽

Bovine Embryo ◽

Sequencing Analysis ◽

Carry Over ◽

Bovine Oocytes

Abstract STUDY QUESTION Does oocyte maturation under lipolytic conditions have detrimental carry-over effects on post-hatching embryo development of good-quality blastocysts after transfer? SUMMARY ANSWER Surviving, morphologically normal blastocysts derived from bovine oocytes that matured under lipotoxic conditions exhibit long-lasting cellular dysfunction at the transcriptomic and metabolic levels, which coincides with retarded post-hatching embryo development. WHAT IS KNOWN ALREADY There is increasing evidence showing that following maturation in pathophysiologically relevant lipotoxic conditions (as in obesity or metabolic syndrome), surviving blastocysts of good (transferable) morphological quality have persistent transcriptomic and epigenetic alteration even when in vitro embryo culture takes place under standard conditions. However, very little is known about subsequent development in the uterus after transfer. STUDY DESIGN, SIZE, DURATION Bovine oocytes were matured in vitro in the presence of pathophysiologically relevant, high non-esterified fatty acid (NEFA) concentrations (HIGH PA), or in basal NEFA concentrations (BASAL) as a physiological control. Eight healthy multiparous non-lactating Holstein cows were used for embryo transfers. Good-quality blastocysts (pools of eight) were transferred per cow, and cows were crossed over for treatments in the next replicate. Embryos were recovered 7 days later and assessed for post-hatching development, phenotypic features and gene expression profile. Blastocysts from solvent-free and NEFA-free maturation (CONTROL) were also tested for comparison. PARTICIPANTS/MATERIALS, SETTING, METHODS Recovered Day 14 embryos were morphologically assessed and dissected into embryonic disk (ED) and extraembryonic tissue (EXT). Samples of EXT were cultured for 24 h to assess cellular metabolic activity (glucose and pyruvate consumption and lactate production) and embryos’ ability to signal for maternal recognition of pregnancy (interferon-τ secretion; IFN-τ). ED and EXT samples were subjected to RNA sequencing to evaluate the genome-wide transcriptome patterns. MAIN RESULTS AND THE ROLE OF CHANCE The embryo recovery rate at Day 14 p.i. was not significantly different among treatment groups (P > 0.1). However, higher proportions of HIGH PA embryos were retarded in growth (in spherical stage) compared to the more elongated tubular stage embryos in the BASAL group (P < 0.05). Focusing on the normally developed tubular embryos in both groups, HIGH PA exposure resulted in altered cellular metabolism and altered transcriptome profile particularly in pathways related to redox-regulating mechanisms, apoptosis, cellular growth, interaction and differentiation, energy metabolism and epigenetic mechanisms, compared to BASAL embryos. Maturation under BASAL conditions did not have any significant effects on post-hatching development and cellular functions compared to CONTROL. LARGE-SCALE DATA The datasets of RNA sequencing analysis are available in the NCBI’s Gene Expression Omnibus (GEO) repository, series accession number GSE127889 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE127889). Datasets of differentially expressed genes and their gene ontology functions are available in the Mendeley datasets at http://dx.doi.org/10.17632/my2z7dvk9j.2. LIMITATIONS, REASONS FOR CAUTION The bovine model was used here to allow non-invasive embryo transfer and post-hatching recovery on Day 14. There are physiological differences in some characteristics of post-hatching embryo development between human and cows, such as embryo elongation and trophoblastic invasion. However, the main carry-over effects of oocyte maturation under lipolytic conditions described here are evident at the cellular level and therefore may also occur during post-hatching development in other species including humans. In addition, post-hatching development was studied here under a healthy uterine environment to focus on carry-over effects originating from the oocyte, whereas additional detrimental effects may be induced by maternal metabolic disorders due to adverse changes in the uterine microenvironment. RNA sequencing results were not verified by qPCR, and no solvent control was included. WIDER IMPLICATIONS OF THE FINDINGS Our observations may increase the awareness of the importance of maternal metabolic stress at the level of the preovulatory oocyte in relation to carry-over effects that may persist in the transferrable embryos. It should further stimulate new research about preventive and protective strategies to optimize maternal metabolic health around conception to maximize embryo viability and thus fertility outcome. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Flemish Research Fund (FWO grant 11L8716N and FWO project 42/FAO10300/6541). The authors declare there are no conflicts of interest.

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Effect of high‐density lipoprotein on oocyte maturation and bovine embryo development in vitro

Reproduction in Domestic Animals ◽

10.1111/rda.13373 ◽

2018 ◽

Vol 54 (3) ◽

pp. 445-455

Author(s):

Joao Alveiro Alvarado Rincón ◽

Jorgea Pradieé ◽

Mariana Härter Remião ◽

Tiago Veiras Collares ◽

Bruna Mion ◽

...

Keyword(s):

High Density Lipoprotein ◽

Embryo Development ◽

Oocyte Maturation ◽

Density Lipoprotein ◽

High Density ◽

Bovine Embryo ◽

Development In Vitro

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Follicular Fluid from Infertile Women with Mild Endometriosis Impairs In Vitro Bovine Embryo Development: Potential Role of Oxidative Stress

Revista Brasileira de Ginecologia e Obstetrícia / RBGO Gynecology and Obstetrics ◽

10.1055/s-0040-1718443 ◽

2021 ◽

Author(s):

Vanessa Silvestre Innocenti Giorgi ◽

Rui Alberto Ferriani ◽

Paula Andrea Navarro

Keyword(s):

Oxidative Stress ◽

Embryo Development ◽

Follicular Fluid ◽

Formation Rate ◽

Fertilization In Vitro ◽

Hatching Rate ◽

Bovine Embryo ◽

Infertile Women ◽

Bovine Oocytes

Abstract Objective To investigate whether follicular fluid (FF) from infertile women with mild endometriosis (ME) alters in vitro bovine embryo development, and whether the antioxidants N-acetyl-cysteine (NAC) and/or L-carnitine (LC) could prevent such damages. Methods Follicular fluid was obtained from infertile women (11 with ME and 11 control). Bovine oocytes were matured in vitro divided in: No-FF, with 1% of FF from control women (CFF) or ME women (MEFF); with 1.5 mM NAC (CFF + NAC, MEFF + NAC), with 0.6 mg/mL LC (CFF + LC, MEFF + LC), or both antioxidants (CFF + NAC + LC, MEFF + NAC + LC). After in vitro fertilization, in vitro embryo culture was performed for 9 days. Results A total of 883 presumptive zygotes were cultured in vitro. No differences were observed in cleavage rate (p = 0.5376) and blastocyst formation rate (p = 0.4249). However, the MEFF group (12.5%) had lower hatching rate than the No-FF (42.1%, p = 0.029) and CFF (42.9%, p = 0.036) groups. Addition of antioxidants in the group with CFF did not alter hatching rate (p ≥ 0.56), and in groups with MEFF, just NAC increased the hatching rate [(MEFF: 12.5% versus MEFF + NAC: 44.4% (p = 0.02); vs MEFF + LC: 18.8% (p = 0.79); versus MEFF + NAC + LC: 30.8% (p = 0.22)]. Conclusion Therefore, FF from infertile women with ME added to medium of in vitro maturation of bovine oocytes impairs hatching rate, and NAC prevented these damages, suggesting involvement of oxidative stress in worst of oocyte and embryo quality of women with ME.

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Pengaruh Penambahan Chorionic Gonadotrophin pada Medium Maturasi terhadap Kemampuan Maturasi, Fertilisasi, dan Perkembangan Embrio secara In Vitro Kambing Peranakan Ettawa (The Effect of Chorionic Gonadotrophin Addition Into Maturation Medium on The Abili

Buletin Peternakan ◽

10.21059/buletinpeternak.v34i1.101 ◽

2012 ◽

Vol 34 (1) ◽

pp. 8

Author(s):

Nurvina Septi Adifa ◽

Pudji Astuti ◽

Diah Tri Widayati

Keyword(s):

Embryo Development ◽

Oocyte Maturation ◽

Chorionic Gonadotrophin ◽

Fertilization In Vitro ◽

Cleavage Rate ◽

Group I ◽

Maturation Medium ◽

Frozen Semen ◽

Significant Difference

This research was conducted to investigate the effect of chorionic gonadotrophin addition into maturation medium on oocyte maturation, fertilization, and embryo development in vitro of Ettawa crossbred. Oocytes were divided into 3 groups, group I: maturation medium without addition of chorionic gonadotrophin (0), group II: 10 μl/10 ml chorionic gonadotrophin was added into maturation medium (1), group III: 20 μl/10 ml chorionic gonadotrophin was added into maturation medium (2). Oocytes were transferred into 50 μl maturation medium, then covered by mineral oil. Oocyte was incubated at 39oC, 5% CO2, 95% humidity for 24 hours for maturation. Matured oocytes were inseminated with frozen semen–thawed concentration 12.5 x 106/ml. Process of fertilization were carried out on incubator 39oC, 5% CO2, 95% humidity for 5 hours. The fertilized oocytes were transferred into 50 μl drop G–1, then incubated at 39oC, 5% CO2, 95% humidity. Embryo development was monitored every 24 hours. Culture medium was changed every 48 hours. G–2 medium used second day after culture. The variables measured involved oocyte maturation, fertilization, and in vitro cleavage rate. The data were analyzed by chi–square, using SPSS 15.0 program. The result showed no significant difference on the percentage of mature oocytes and fertilization rate were 78.0%, 72.8%, 75.0% and 76.6%, 74.5%, 77.8% respectively. But cleavage rate showed significant difference (P≤0.05) with the values of 40.8%, 11.4%, and 12.2% respectively. Based on the result it could be concluded that chorionic gonadotrophin addition into maturation medium had not increased ettawa crossbred oocytes maturation, fertilization, and in vitro cleavage rate. The best maturation, fertilization, and in vitro cleavage rate were found using maturation medium without any addition of chorionic gonadotrophin.(Key words: Does oocyte, Chorionic gonadotrophin, In vitro maturation, In vitro fertilization, In vitro embryo development)

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