scholarly journals Effects of the donor factors and freezing protocols on the bovine embryonic lipid profile

2021 ◽  
Author(s):  
Sarah Janati Idrissi ◽  
Daniel Le Bourhis ◽  
Antoine Lefevre ◽  
Patrick Emond ◽  
Laurene Le Berre ◽  
...  
Keyword(s):  
Lipid Profile ◽  
High Resolution Mass Spectrometry ◽  
Culture Conditions ◽  
Slow Freezing ◽  
Bovine Embryo ◽  
Lactating Cows ◽  
Physiological Stage ◽  
Donor Factors ◽  
Donor Cows

Abstract Embryo lipid profile is affected by in vitro culture conditions, that lead to an increase in lipids. Efforts have been made to optimize embryo lipid composition as it is associated with their quality. The objective of this study was to evaluate whether the diet supplementation of donor cows (n-3 or n-6 PUFA), or the slow freezing protocols (ethylene glycol sucrose EG-S vs. glycerol trehalose GLY-TRE), or the physiological stage of the donor (nulliparous heifers vs. primiparous lactating cows) may impact the bovine embryo lipid profile. Lipid extracts of 97 embryos were individually analysed by liquid chromatography-high resolution mass spectrometry, highlighting 246 lipids including 85% being overabundant in cow embryos compared to heifer embryos. Among 105 differential lipids, 72 were overabundant after EG-S protocol, including a single glycerophosphate PA(32:1) representing 27.3% of the significantly modulated lipids, suggesting that it is degraded when GLY-TRE is used. No lipids were different according to the n-3 or n-6 supplementation of the donor cows. In conclusion, the embryonic lipid profile was mainly affected by the physiological stage of the donors and the slow freezing protocols. The overabundance of lipids in lactating cow embryos and the resulting lower quality of these embryos is consistent with the lower pregnancy rate observed in cows compared to heifers. Unlike GLY-TRE protocol, EG-S freezing allowed to preserve glycerophospholipids potentially improving the slow freezing of in vitro-produced embryos. Further studies are required to modulate embryo quality and freezability by modulating the lipidome and integrating all stages of embryonic production.

scholarly journals Lipid profile of bovine grade-1 blastocysts produced either in vivo or in vitro before and after slow freezing process

2021 ◽  
Author(s):  
Sarah Janati Idrissi ◽  
Daniel Le Bourhis ◽  
Antoine Lefevre ◽  
Patrick Emond ◽  
Laurene Le Berre ◽  
...  
Keyword(s):  
Lipid Profile ◽  
High Resolution Mass Spectrometry ◽  
High Sensitivity ◽  
Culture Conditions ◽  
Slow Freezing ◽  
Freezing Process ◽  
Before And After ◽  
Lipid Enrichment

Abstract Currently, in vitro embryo production (IVP) is successfully commercially applied in cattle. However, the high sensitivity of embryos to cryopreservation in comparison to in vivo (IVD) embryos slows the dissemination of this biotechnology. Reduced cryotolerance is frequently associated with lipid accumulation in the cytoplasm mainly due to in vitro culture conditions. The objective of this study was to evaluate the lipid composition of biopsied and sexed embryos, produced either in vivo or in vitro from the same Holstein heifers before and after a slow freezing protocol. Lipid extracts were analysed by liquid chromatography-high resolution mass spectrometry, which enabled the detection of 496 features. Our results highlighted a lipid enrichment of IVP embryos in triglycerides and oxidised glycerophospholipids and a reduced abundance in glycerophospholipids. The slow freezing process affected the lipid profiles of IVP and IVD embryos similarly. Lysophosphatidylcholine content was reduced when embryos were frozen/thawed. In conclusion, the embryonic lipid profile is impacted by IVP and slow freezing protocols but not by sex. Lysophosphatidylcholine seemed highly sensitive to cryopreservation and might contribute to explain the lower quality of frozen embryos. Further studies are required to improve embryo freezability by modulating the lipidome.

scholarly journals Lipid profile of bovine grade-1 blastocysts produced either in vivo or in vitro before and after slow freezing process

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sarah Janati Idrissi ◽  
Daniel Le Bourhis ◽  
Antoine Lefevre ◽  
Patrick Emond ◽  
Laurene Le Berre ◽  
...  
Keyword(s):  
Lipid Profile ◽  
High Resolution Mass Spectrometry ◽  
High Sensitivity ◽  
Culture Conditions ◽  
Slow Freezing ◽  
Freezing Process ◽  
Before And After ◽  
Lipid Enrichment

AbstractCurrently, in vitro embryo production (IVP) is successfully commercially applied in cattle. However, the high sensitivity of embryos to cryopreservation in comparison to in vivo (IVD) embryos slows the dissemination of this biotechnology. Reduced cryotolerance is frequently associated with lipid accumulation in the cytoplasm mainly due to in vitro culture conditions. The objective of this study was to evaluate the lipid composition of biopsied and sexed embryos, produced either in vivo or in vitro from the same Holstein heifers before and after a slow freezing protocol. Lipid extracts were analysed by liquid chromatography-high resolution mass spectrometry, which enabled the detection of 496 features. Our results highlighted a lipid enrichment of IVP embryos in triglycerides and oxidised glycerophospholipids and a reduced abundance in glycerophospholipids. The slow freezing process affected the lipid profiles of IVP and IVD embryos similarly. Lysophosphatidylcholine content was reduced when embryos were frozen/thawed. In conclusion, the embryonic lipid profile is impacted by IVP and slow freezing protocols but not by sex. Lysophosphatidylcholine seemed highly sensitive to cryopreservation and might contribute to explain the lower quality of frozen embryos. Further studies are required to improve embryo freezability by modulating the lipidome.

scholarly journals Lipid Profiling of Peri-implantation Endometrium in Patients With Premature Progesterone Rise in the Late Follicular Phase

2019 ◽  
Vol 104 (11) ◽  
pp. 5555-5565 ◽  
Author(s):  
Jingjie Li ◽  
Yue Gao ◽  
Lihuan Guan ◽  
Huizhen Zhang ◽  
Pan Chen ◽  
...  
Keyword(s):  
Lipid Profile ◽  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
High Resolution Mass Spectrometry ◽  
Follicular Phase ◽  
Lipid Profiling ◽  
Male Factor ◽  
Serum Progesterone ◽  
Late Follicular Phase

Abstract Context Late follicular phase elevation in serum progesterone (P) during controlled ovarian hyperstimulation negatively affects the outcome of assisted reproductive technology by contributing to endometrial-embryo asynchrony. There are still no data on lipid metabolite alterations during this process. Objectives To investigate alterations in the lipid profile during the window of implantation in patients with premature P rise. Design Lipidomic variations in the endometrium were evaluated by ultrahigh-performance liquid chromatography coupled with electrospray ionization high-resolution mass spectrometry. Setting University assisted reproductive medicine unit. Patients or Other Participants Forty-three patients undergoing in vitro fertilization/intracytoplasmic sperm injection because of a tubal factor or male factor infertility were included in this study. The patients were divided into a high P group (P ≥ 1.5 ng/mL, 15 patients) and a normal P group (P < 1.5 ng/mL, 28 patients) on the day of human chorionic gonadotropin administration. Interventions The endometrial tissues were obtained by Pipelle biopsy 7 days after human chorionic gonadotropin administration. Main Outcome Measures Alterations in lipid metabolites. Results A total of 1026 ions were identified, and 25 lipids were significantly upregulated. The endometrial lipid profile was characterized by substantial increases in the concentrations of phosphatidylcholine, phosphatidylethanolamine, lysophosphatidylcholine, diacylglycerol, ceramide, phosphatidylinositol, and phosphatidylserine in patients with a premature P rise at the end of the follicular phase. The correlation analysis between P levels and lipids showed a stronger negative correlation between phosphatidylethanolamine or phosphatidylserine and P levels. Conclusions Premature P elevation disrupts the lipid homeostasis of the endometrium during the peri-implantation period. The altered lipid levels may impair endometrial receptivity and early embryo implantation.

Effects of different cryopreservation methods on post-thaw culture conditions of in vitro produced bovine embryos

Zygote ◽  
2011 ◽  
Vol 20 (2) ◽  
pp. 117-122 ◽  
Author(s):  
Alessandra Corallo Nicacio ◽  
Renata Simões ◽  
Fabiola Freitas de Paula-Lopes ◽  
Flavia Regina Oliveira de Barros ◽  
Maria Angelica Peres ◽  
...  
Keyword(s):  
Culture Conditions ◽  
Slow Freezing ◽  
Control Group ◽  
Hatching Rate ◽  
Rapid Freezing ◽  
Bovine Embryos ◽  
Group 10 ◽  
Controlled Freezing ◽  
Hatching Rates

SummaryThe aim of this work was to evaluate the effect of cryopreservation protocols on subsequent development of in vitro produced bovine embryos under different culture conditions. Expanded in vitro produced blastocysts (n = 600) harvested on days 7–9 were submitted to controlled freezing [slow freezing group: 10% ethylene glycol (EG) for 10 min and 1.2°C/min cryopreservation]; quick-freezing [rapid freezing group: 10% EG for 10 min, 20% EG + 20% glycerol (Gly) for 30 s]; or vitrification [vitrification group: 10% EG for 10 min, 25% EG + 25% Gly for 30 s] protocols. Control group embryos were not exposed to cryoprotectant or cryopreservation protocols and the hatching rate was evaluated on day 12 post-insemination. In order to evaluate development, frozen–thawed embryos were subjected to granulosa cell co-culture in TCM199 or SOFaa for 4 days. Data were analyzed by PROC MIXED model using SAS Systems for Windows®. Values were significant at p < 0.05. The hatching rate of the control group was 46.09%. In embryos cultured in TCM199, slow freezing and vitrification group hatching rates were 44.65 ± 5.94% and 9.43 ± 6.77%, respectively. In embryos cultured in SOFaa, slow freezing and vitrification groups showed hatching rates of 11.65 ± 3.37 and 8.67 ± 4.47%, respectively. In contrast, the rapid freezing group embryos did not hatch, regardless of culture medium. The slow freezing group showed higher hatching rates than other cryopreservation groups. Under such conditions, controlled freezing (1.2°C/min) can be an alternative to cryopreservation of in vitro produced bovine embryos.

scholarly journals Cryopreservation of in vitro-produced bovine embryos by vitrification: In pursuit of a simplified, standardized procedure that improves pregnancy rates to promote cattle industry use

2020 ◽  
Vol 36 (3) ◽  
pp. 251-270
Author(s):  
Van Do ◽  
Andrew Taylor-Robinson
Keyword(s):  
Slow Rate ◽  
Survival Rates ◽  
Slow Freezing ◽  
Embryo Cryopreservation ◽  
Human Embryos ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Cattle Industry

The goal of cryopreservation is to retain the original stage of gametes and embryos after they have endured cooling and warming. Slow freezing is a standard method for in vivo-derived bovine embryo cryopreservation, threefifths of such embryos being frozen by this method globally. However, it is evident that slow freezing is not efficient for cryopreserving in vitro-produced bovine embryos. Hence, only one-third of in vitro-produced bovine embryos are cryopreserved. Vitrification is a preferred method for storage of human embryos; consequently, it has been explored as a novel means to store in vitro-produced bovine embryos, for which it shows considerable promise as an alternative to slow freezing. This is due to several reasons: vitrification is often less time-consuming than slow freezing; it does not need expensive slow rate freezing machines; and it has been proven to have comparatively higher survival rates. Yet, in the cattle industry vitrification continues to present shortcomings, such as possible toxicity of vitrification solutions and failure to standardize methods, which pose a challenge for its application to in vitro-produced bovine embryos. Therefore, determining the most suitable procedure is crucial to make vitrification more practical in commercial settings.

126 A Comparison of Two Different Follicular Coasting Periods for In Vitro Embryo Production in Indian Nelore Cattle

2018 ◽  
Vol 30 (1) ◽  
pp. 203
Author(s):  
P. Tiwari ◽  
S. Zawar ◽  
J. H. Pryor ◽  
C. R. Looney ◽  
R. Kaushik ◽  
...  
Keyword(s):  
Animal Health ◽  
Bos Indicus ◽  
Embryo Production ◽  
Treatment Groups ◽  
Lactating Cows ◽  
Oocyte Recovery ◽  
Minimum Interval ◽  
Nelore Cattle ◽  
Donor Cows

Ongole, also known as Nelore (Bos indicus) cattle, are indigenous to the Andhra region in the Prakasam District in the State of Andhra Pradesh in India. A better understanding and utilisation of follicular wave dynamics within this breed would ultimately enhance oocyte and potential embryo production. Therefore, the aim of this study was to evaluate the differences between coasting periods of 24 h (S1) and 36 h (S2) on oocyte recovery, the rate of viable oocytes, cleavage, and Day 7 blastocyst rates of Nelore cattle in India. A total of 58 ovum pick-up (OPU) sessions (29 per treatment) were performed on 32 healthy donor cows that were randomly assigned to 1 of 2 coasting treatments (S1 or S2). Donors were stimulated as follows: 2.5 mL of gonadotropin-releasing hormone (GnRH; Receptal, MSD Animal Health, New Zealand) given IM on Day 1 followed by once-daily descending dose of Folltropin® (FSH, Vetoquinol, Canada) on Days 3 to 5 for a total of 180 mg. Cumulus-oocyte complexes were collected following OPU on Day 6 either at 24 (S1) or 36 h (S2) following the last FSH injection (coasting period). Donors were subject to OPU 1 to 3 times with a minimum interval of 15 days between procedures from March to April 2017. All 32 donor cows were non-lactating at the time of aspiration and divided equally between treatment groups. A total of 1492 follicles produced 850 total oocytes with oocyte recovery numbers for treatments S1 and S2 (785, 707; 441, 409; respectively). All data were analysed by ANOVA (P < 0.05). The mean number of follicles aspirated from S1 (27 ± 20.2) was not significantly different from that of S2 (24.4 ± 14.4). For S1, 393/441 (89%) quality oocytes were utilised for culture compared with 323/409 (78.9%) for S2, with no differences between rates. Additionally, there were no differences between mean number of oocytes, cleaved embryos, and blastocysts for S1 (15.2 ± 12.7; 9.9 ± 9.2; 4.3 ± 5.4) and S2 (14.1 ± 10; 7.4 ± 6.0; 3.6 ± 3.3; respectively). In conclusion, there were no differences found between 24- or 36-h coasting periods of Nelore cattle undergoing OPU for follicle counts, oocyte recovery, viable oocyte rates, cleavage, and blastocyst rates. Further research is needed to determine whether different stimulation protocols, the use of lactating cows, or coasting periods could alter outcomes.

scholarly journals Embryo transfer as a tool for improving fertility of heat-stressed dairy cattle

2007 ◽  
Author(s):  
Peter J. Hansen ◽  
Amir Arav
Keyword(s):  
Elevated Temperature ◽  
Heat Shock ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Caspase 3 ◽  
Culture Conditions ◽  
Caspase Activity ◽  
Lactating Cows ◽  
Selection Of

The overall objective of the current proposal is to develop procedures to improve the pregnancy rate achieved following transfer of fresh or cryopreserved embryos produced in the laboratory into heat-stress recipients. The overall hypothesis is that pregnancy rate in heat-stressed lactating cows can be improved by use of embryo transfer and that additional gains in pregnancy rate can be achieved through development of procedures to cryopreserve embryos, select embryos most likely to establish and maintain pregnancy after transfer, and to enhance embryo competence for post-transfer survival through manipulation of culture conditions. The original specific objectives were to 1) optimize procedures for cryopreservation (Israel/US), 2) develop procedures for identifying embryos with the greatest potential for development and survival using the remote monitoring system called EmbryoGuard (Israel), 3) perform field trials to test the efficacy of cryopreservation and the EmbryoGuard selection system for improving pregnancy rates in heat-stressed, lactating cows (US/Israel), 4) test whether selection of fresh or frozen-thawed blastocysts based on measurement of group II caspase activity is an effective means of increasing survival after cryopreservation and post-transfer pregnancy rate (US), and 5) identify genes in blastocysts induced by insulin-like growth factor-1 (IGF-1) (US). In addition to these objectives, additional work was carried out to determine additional cellular determinants of embryonic resistance to heat shock. There were several major achievements. Results of one experiment indicated that survival of embryos to freezing could be improved by treating embryos with cytochalasin B to disrupt the cytoskeleton. An additional improvement in the efficacy of embryo transfer for achieving pregnancy in heat-stressed cows follows from the finding that IGF-1 can improve post-transfer survival of in vitro produced embryos in the summer but not winter. Expression of several genes in the blastocyst was regulated by IGF-1 including IGF binding protein-3, desmocollin II, Na/K ATPase, Bax, heat shock protein 70 and IGF-1 receptor. These genes are likely candidates 1) for developing assays for selection of embryos for transfer and 2) as marker genes for improving culture conditions for embryo production. The fact that IGF-1 improved survival of embryos in heat-stressed recipients only is consistent with the hypothesis that IGF-1 confers cellular thermotolerance to bovine embryos. Other experiments confirmed this action of IGF-1. One action of IGF-1, the ability to block heat-shock induced apoptosis, was shown to be mediated through activation of the phosphatidylinositol 3-kinase pathway. Other cellular determinants of resistance of embryos to elevated temperature were identified including redox status of the embryo and the ceramide signaling pathway. Developmental changes in embryonic apoptosis responses in response to heat shock were described and found to include alterations in the capacity of the embryo to undergo caspase-9 and caspase-3 activation as well as events downstream from caspase-3 activation. With the exception of IGF-1, other possible treatments to improve pregnancy rate to embryo transfer were not effective including selection of embryos for caspase activity, treatment of recipients with GnRH.and bilateral transfer of twin embryos. In conclusion, accomplishments achieved during the grant period have resulted in methods for improving post-transfer survival of in vitro produced embryos transferred into heat-stressed cows and have lead to additional avenues for research to increase embryo resistance to elevated temperature and improve survival to cryopreservation. In addition, embryo transfer of vitrified IVF embryos increased significantly the pregnancy rate in repeated breeder cows.

70 ONE-STEP CRYOPROTECTANT DILUTION FOLLOWING VITRIFICATION OF IN VITRO-PRODUCED BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 182
Author(s):  
R. Morató ◽  
T. Mogas
Keyword(s):  
Embryo Transfer ◽  
Statistical Significance ◽  
Survival Rates ◽  
Blastocyst Stage ◽  
Slow Freezing ◽  
Embryo Cryopreservation ◽  
Dilution Method ◽  
Bovine Embryo ◽  
One Step

Although slow freezing continues to be the most widely used technique of cryopreservation for bovine in vivo- and in vitro-produced embryos, vitrification has been tested in different species with good results, especially when dealing with in vitro-produced embryos. Vitrification represents a minor expense in time and equipment associated with cryopreservation compared with conventional slow freezing. However, vitrification, which is the most common method for human embryo cryopreservation, has not been widely adopted by embryo-transfer practitioners for commercial use in cattle. In general, vitrification requires gradual cryoprotectant dilution in a laboratory setting, and it is difficult to perform in the field. The objective of this study was to develop a one-step dilution method suitable for one-step bovine embryo transfer using the cryotop vitrification method. Embryos produced in vitro by standard procedures were vitrified at the blastocyst stage at Day 7 post-insemination in a mixture of 15% ethylene glycol + 15% dimethyl sulfoxide + 0.5 M sucrose using cryotop devices. Embryos were randomly assigned to 1 of 3 warming methods: (1) W3: warming was carried out following the cryotop method (1 M sucrose for 1 min, 0.5 M sucrose for 3 min, and 0 M sucrose for 6 min); (2) W1/0.5: embryos were warmed directly in 0.5 M sucrose for 3 min; and (3) W1/0: embryos were warmed directly in 0 M sucrose for 5 min. Survival rates were assessed in terms of blastocyst re-expansion, hatching, and hatched status at 3 and 24 h after warming. Data were analyzed using the statistical analysis systems package (SAS, v9.1). Data from at least 3 replicates were collected. Comparisons of vitrified–warmed blastocyst survival rates between groups were performed using the chi-squared test. The level of statistical significance was set at P < 0.05. When embryo survival was evaluated at 3 h postwarming, embryos warmed using the 3-step dilution protocol and those warmed directly in 0.5 M sucrose showed higher percentages of survival (W3: 89.8%, n = 98; W1/0.5: 87.5%, n = 64; P < 0.05) than those blastocysts that were warmed directly in 0 M sucrose (W1/0: 66.4%, n = 146). However, similar rates irrespective of the warming procedure were observed at 24 h postwarming (W3: 85.7%, W1/0.5: 88.2%, W1/0: 70.5%). Warmed in vitro-produced embryos exposed to W3 (47.6%) and W1/0.5 (35.6%) achieved higher percentages of embryos developing to the hatched blastocyst stage after 24 h of culture than those embryos warmed in W1/0 (20.4%; P < 0.05). Our results indicate that direct warming and dilution of cyotop-vitrified embryos in 0.5 M sucrose for 3 min may enable one-step bovine embryo transfer without requirement of a microscope or other laboratory equipment, simplifying the embryo-transfer procedure of vitrified embryos on farm at the same level of complexity as carrying out AI. Support came from Spanish MEC (RZ2010-00015-0-00; AGL2010-19069) and Generalitat de Catalunya (2009 SGR 621).

128 CULTURE CONDITIONS AFFECT THE SEX RATIO OF IN VITRO PRODUCED BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 211
Author(s):  
M. De Blasi ◽  
M. Rubessa ◽  
G. Zullo ◽  
L. Boccia ◽  
V. Longobardi ◽  
...  
Keyword(s):  
Energy Source ◽  
Sex Ratio ◽  
Trisodium Citrate ◽  
Culture Conditions ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Chi Square ◽  
Group A ◽  
Group B

Most systems for producing bovine embryos in vitro use glucose as an energy source despite putative toxic effects. Glucose has a selective embryotoxicity towards female embryos, due to the higher expression of the X-linked glucose-6-phosphate dehydrogenase gene (Kimura et al. 2005 Mol. Reprod. Dev. 72, 201–207). Recently, the replacement of glucose with citrate and myo-inositol in SOF medium supplemented with 5% bovine serum (BS) increased the percentage of female embryos (Rubessa et al. 2011 Theriogenology 76, 1347–1355). Serum also affects the sex ratio of in vitro-produced (IVP) bovine embryos, favoring the male gender (Gutierrez-Adan et al. 2001 Theriogenology 55, 1117–1126). The aim of this work was to evaluate the effect of glucose replacement with myo-inositol during in vitro culture, in the presence of either BS or BSA, on bovine embryo sex ratio. Abattoir-derived oocytes (n = 1164, over 4 replicates) were matured and fertilized in vitro as previously described (Rubessa et al. 2011). After 20 to 22 h of gametes co-incubation, zygotes were denuded and cultured for 7 days in SOF with: group A) 0.34 mM trisodium citrate + 2.77 mM myo-inositol + 5% BS (n = 287); group B) 0.34 mM tri-sodium citrate + 2.77 mM myo-inositol + 8 mg mL–1 BSA(n = 290); group C) 1.5 mM glucose + 5% BS (n = 302) and group D) 1.5 mM glucose + 8 mg mL–1 BSA (n = 285). Representative samples of blastocysts produced in each group (n = 96, 58, 99, and 70, respectively in groups A, B, C, and D) were sexed by PCR as previously described (Rubessa et al. 2011). Differences among groups in blastocyst yields were analyzed by ANOVA. The percentages of female embryos were analyzed by chi-square test. Blastocyst rates in group C were lower (28.1%) than those recorded in groups A, B, and D (35.9, 41.0 and 36.1%, respectively; P < 0.01). A higher (P < 0.05) percentage of female embryos was observed in group A (61.5%) compared to group C (45.5%), with intermediate values in groups B (51.7%) and D (60.0%). Therefore, the replacement of glucose with citrate and myo-inositol favored the development of female embryos in the presence of BS but was ineffective in the presence of BSA. Furthermore, when glucose was the energy source, a tendency to greater incidence of female embryos was observed when the medium was supplemented with BSA rather than BS (P = 0.06). As a small amount of glucose is present in the BS, we hypothesize an additional glucose-dependent toxic effect on female embryos in group C. However, we cannot rule out that other factors present in the BS may interact with the energy source, playing a role in determining the sex ratio. Furthermore, the shift in sex ratio in favor of males or females embryo can be due to a better development of embryo of one sex, or to the delayed development or degeneration of embryos of the other sex. In conclusion, these results suggest that manipulating the metabolic profile of the embryos during culture may have an impact on both blastocyst production and sex ratio.

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