141 SUPERIORITY OF FEMALE EMBRYO PRODUCTION SYSTEM BY IN VIVO-MATURED OOCYTE AND X-SORTED SPERM IN BROWN SWISS COWS

2014 ◽  
Vol 26 (1) ◽  
pp. 184
Author(s):  
T. Yamanouchi ◽  
H. Matsuda ◽  
M. Ohtake ◽  
K. Masaki ◽  
E. Horiguchi ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Female Embryo ◽  
Brown Swiss ◽  
Embryo Recovery ◽  
Matured Oocyte ◽  
Matured Groups

Embryo transfer using a female embryo is an effective tool for offspring production on dairy industry; however, embryo production by embryo recovery (ER) using X-sorted semen is not sufficient because non-fertilized oocytes are recovered frequently. In Holstein cows, we developed a system for high blastocyst production that was performed by IVF using X-sorted sperm and in vivo-matured oocytes obtained by ovum pickup (OPU) after superstimulation. The purpose of this study was to adjust this system to Brown Swiss cows, comparing between ER and embryo production from oocytes derived from OPU with or without superstimulation. In the ER group, cows (n = 10) received a CIDR (Day 0) and 2 mg of oestradiol-benzoate on Day 1. A total of 30 Armour Units of FSH were injected into cows twice a day, with decreasing doses from the evening of Day 5 to the morning of Day 9. On the evening of Day 7 or 8, 0.75 mg of prostaglandin F2α (cloprostenol) was injected. The CIDR was removed on Day 8 or 9 and 0.2 mg of gonadotropin-releasing hormone (GnRH; fertirelin acetate) were injected on Day 9 or 10. At oestrus, AI was carried out twice, 12 h apart, with a total of 4 straws of X-sorted semen per cow. In the OPU group, cows (n = 7) were subjected to OPU without any pretreatment, collected immature oocytes were in vivo matured for 20 to 22 h, followed by IVF using X-sorted sperm for 6 h; then, presumptive zygotes were in vitro cultured (IVC) for 9 days. In the in vivo-matured oocyte group (matured group), a CIDR was inserted (Day 0) in cows (n = 4), all follicles larger than 8 mm were removed on Day 5. Administration of FSH, prostaglandin F2α, and GnRH, as well as withdrawal of CIDR, were performed as in the ER group. In vivo-matured oocytes were collected from follicles larger than 5 mm by OPU at 25 to 26 h following GnRH injection; collected oocytes with expanded cumulus cells were fertilized with X-sorted sperm 30 h after GnRH. After 6 h of IVF, presumptive zygotes were transferred to in vitro culture, as in the OPU group. Data were compared among 3 groups; the ER group was analysed for number of CL, collected embryos, and normal embryos, against the number of aspirated follicles, collected oocytes used for IVF, and formed blastocysts in the OPU and matured groups, respectively, by Tukey-kramer test after ANOVA. There were no differences between the number of CL in the ER group and the number of follicles in the OPU and matured groups (16.4 ± 5.3 v. 31.6 ± 22.7 v. 18.5 ± 4.7, mean ± s.d., respectively). Also the number of collected embryos in the ER group and number of oocytes for IVF in the OPU and matured groups (12.8 ± 7.6 v. 14.9 ± 11.8 v. 17.8 ± 7.7, respectively) was similar. However, the number of blastocysts in the matured group (13.0 ± 5.9; P < 0.01) was higher than that in the OPU group (3.0 ± 2.2) and in the ER group (2.8 ± 3.7). For female embryo production in Brown Swiss cows using X-sorted sperm, the system of IVF with in vivo matured oocytes obtained by OPU is more effective than ER or OPU without pretreatment.

164 The Origin of Oocyte, In Vitro-Matured Oocyte With/Without Super-Stimulation, and In Vivo-Matured Oocyte Influence the Timing of Cleavage in Early Embryo and Oxygen Consumption of Blastocyst After IVF in Japanese Black Cow

2018 ◽  
Vol 30 (1) ◽  
pp. 221
Author(s):  
T. Yamanouchi ◽  
H. Matsuda ◽  
M. Ohtake ◽  
Y. Ogata ◽  
Y. Aikawa ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Oxygen Consumption ◽  
Prostaglandin F2α ◽  
Early Embryo ◽  
Developmental Competence ◽  
Induction Treatment ◽  
Time Interval ◽  
Matured Oocyte

It has been reported that in vitro- and in vivo-matured oocyte obtained from fully growth follicles have high developmental competence. Furthermore, the timing of cleavage in early embryo after IVF affect pregnancy success after embryo transfer. It is still unknown whether origin of oocyte affects the timing of cleavage. In this study, we examined the influence of oocyte origin on cleavage timing of early embryo after IVF. Japanese Black cows were used as donors. Oocytes derived from non-stimulation follicles (control: CON), fully grown follicles after super-stimulation treatment (SST) and follicles just before ovulation after ovulation-induction treatment (in vivo-matured oocyte: VIVO) were obtained by ovum pick-up (OPU). In the CON group, OPU was conducted on arbitrary days except oestrus. In SST group, dominant follicles were aspirated and a CIDR was inserted into the vagina on Day 0, and then FSH was injected twice a day from the evening of Day 1 to the morning of Day 5 with decreasing doses in total 20 AU. In the evening of Day 4, prostaglandin F2α (0.5 mg of cloprostenol) was administered. On Day 6, SST oocytes were collected after CIDR withdrawl. In the VIVO group, the treatment was carried out as SST until prostaglandin F2α administration, and then CIDR withdrawal and administration of gonadotropin-releasing hormone (GnRH, 0.2 mg of fertirelin acetate) performed on the evening of Day 4 and morning of Day 5, respectively. The VIVO oocytes were collected at 25 to 26 h after GnRH. The CON and SST oocytes were inseminated after 20 to 22 h of IVM, and VIVO oocytes were inseminated at 30 h after GnRH, with 3 × 106 sperm mL−1, respectively. After 6 h of IVF, presumptive zygotes were individually cultured for 168 h, using a well-of-the-well dish (Dai-Nippon-Print, Japan) and were observed by time-lapse cinematography (CCM-4MZS; Astec, Japan) to analyse the cleavage timing of embryos. Oxygen consumption (O2) was measured in blastocysts on 168 hpi with a scaning electrochemical microscopy system (HV-405SP; Hokuto Denko, Japan). Statistical analysis was carried out by Steel-Dwass test for the timing of cleavage and Tukey-Kramer test for O2. In CON (n = 15), SST (n = 25), and VIVO (n = 36), the time of first cleavage was 27.5, 29.1, and 26.1 hpi, that of second cleavage was 38.9, 40.3, and 36.0 hpi, and that of third cleavage was 48.5, 46.1, and 45.9 hpi, respectively. These cleavage times were shorter in VIVO than in CON and SST (P < 0.01). The time interval between first and second cleavage (2nd cell cycle) was shorter in VIVO (10.1; P < 0.01) than CON (11.4) and SST (11.2). The time interval between second and third (3rd cell cycle) were shorter (P < 0.01) in SST (9.4) than in VIVO (10.1), and in VIVO than in CON (10.2), respectively. Consumption of O2 was lower (P < 0.01) in CON (0.61 × 10−14 mol s−1) than in SST (0.94 × 10−14 mol s−1) and VIVO (0.94 × 10−14 mol s−1). These results suggest that the origin of oocyte influences the length of cell cycle and O2 consumption of blastocyst producted in vitro.

134 EFFECT OF ELEVATED CIRCULATING PROGESTERONE CONCENTRATION ON DEVELOPMENT OF IN VITRO PRODUCED BOVINE ZYGOTES IN VIVO

2008 ◽  
Vol 20 (1) ◽  
pp. 147 ◽  
Author(s):  
F. Rings ◽  
F. Carter ◽  
M. Hölker ◽  
A. Kuzmany ◽  
U. Besenfelder ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Blastocyst Stage ◽  
Early Embryo ◽  
Progesterone Concentration ◽  
Interferon Tau ◽  
Endoscopic Techniques ◽  
Embryo Recovery ◽  
Chi Square Analysis

Elevated concentrations of circulating progesterone in the immediate post-conception period have been associated with an increase in embryonic growth rate, interferon-tau production, and pregnancy rate in cattle and sheep. Much of this effect is likely mediated via downstream effects of progesterone-induced changes in gene expression in the tissues of the uterus. However, whether or not progesterone has a direct effect on the embryo also is unknown and, at least in vivo, in single ovulating animals, is difficult to assess. Using state-of-the-art endoscopic techniques, the objective of this study was to examine the effect of elevated progesterone on the development of IVP zygotes transferred to the oviducts of cattle with high or normal circulating progesterone concentrations. Simmental heifers (n = 14) were synchronized using a combination of 2 injections of a prostaglandin F2α analogue administered 11 days apart and gonadotropin-releasing hormone. Only animals exhibiting a clear standing oestrus (= day 0) were used. In order to produce animals with divergent progesterone concentrations, half of the animals received a PRID on day 3 of the oestrous cycle, which was left in place until embryo recovery. All animals were blood sampled daily from days 0 to 7. Cleaved embryos were transferred using endoscopy to the ipsilateral oviduct of each recipient on day 2 and recovered by non-surgically flushing the oviduct and the uterus on day 7. The number of embryos developing to the morula/blastocyst stage was recorded at recovery and following overnight culture in CR1aa medium. Data were analyzed by chi-square analysis. Insertion of a PRID on day 3 resulted in a significant elevation in progesterone concentrations from day 4 (2.36 ± 0.16 ng mL–1 v. 0.54 ± 0.10 ng mL–1, P < 0.001) until day 6 (1.98 ± 0.22 ng mL–1 v. 0.95 ± 0.17 ng mL–1; P < 0.01). The recovery rate was lower in animals that received a PRID (P < 0.05). However, there was no effect of progesterone on the proportion of embryos developing to the morula/blastocyst stage. These results suggest that elevated concentrations of progesterone do not affect the ability of the early embryo to reach the blastocyst stage in vivo and that the reported positive effect of high progesterone levels in terms of fertility are manifested after day 8. Table 1. Effect of elevated progesterone concentration on development of in vitro produced bovine zygotes in vivo

240 EFFECTS OF REPEATED SUPEROVULATION ON EMBRYO YIELDS IN SOME TURKISH NATIVE GOAT BREEDS

2013 ◽  
Vol 25 (1) ◽  
pp. 268
Author(s):  
M. Kaymaz ◽  
A. R. Agaoglu ◽  
K. Karakas ◽  
I. Pir Yagci ◽  
O. Korkmaz Agaoglu ◽  
...  
Keyword(s):  
Prostaglandin F2α ◽  
Repeated Administration ◽  
Corpora Lutea ◽  
Embryo Production ◽  
Recovery Rates ◽  
Embryo Recovery ◽  
The Third ◽  
Friedman's Test ◽  
Negative Effect

The Angora, Kilis, Honamli, Hair, and Norduz are native goat breeds in Turkey and are currently in danger of extinction. This study aimed to assess the efficacy of the repeated administration of a superovulatory (SOV) protocol for in vivo embryo production in these breeds. A total of 14 Angora, 15 Kilis, 10 Honamli, 10 Hair, and 9 Norduz goats were used in this work. The synchronization procedure was started on Day 5 after visible oestrus by using controlled internal drug release dispensers (CIDR®) for 11 days. Administration of FSH (Folltropin®) began on Day 9 (twice daily) and continued for 3 days (total dose: 200 mg; 50 mg × 2.30 mg × 2.20 mg × 2). A dose of prostaglandin F2α (1.6 mg; Dalmazin®) was injected together with first FSH injection. Gonadotropin-releasing hormone (Receptal®: 100 µg) was injected 6 h before mating. All goats in oestrus were naturally mated twice a day for 3 days. Ovarian examination (number of corpora lutea) and embryo recovery were performed by laparotomy on Day 6 after CIDR® withdrawal. Each uterine horn was flushed, and the embryos were recovered and counted. To avoid intra-abdominal adhesions, a 2.5% heparin solution was used during flushing. The SOV procedure was repeated once per year during the breeding season (2009 to 2011). Fertilization and recovery rates were calculated. Differences in the SOV response and embryo yields were evaluated by Friedman’s test. In Hair goats, the number of corpora lutea decreased significantly (P < 0.05) during the third SOV cycle (12.7 ± 6.2, 14.0 ± 9.1, and 6.8 ± 5.6, respectively, for cycles 1, 2, and 3), whereas no effect of the cycle was observed in the remainder of breeds. The number of expanded blastocysts increased considerably during the third cycle in Angora (0, 0.2 ± 0.8, and 1.4 ± 2.9, respectively, for cycles 1, 2, and 3), Kilis (0.2 ± 0.4, 0.3 ± 1.3, and 4.2 ± 5.0), and Honamli (0, 1.3 ± 1.7, and 3.6 ± 4.5) goats, whereas a significant decrease was observed in Norduz goats (2.4 ± 5.0, 1.8 ± 2.0, and 0.1 ± 0.3; P < 0.05). The mean numbers of unfertilized oocytes were found to be significantly increased in Angora (0.4 ± 1.6, 0, and 2.1 ± 4.1, respectively, for cycles 1, 2, and 3), Kilis (0, 1.3 ± 3.9, and 3.1 ± 5.2), and Honamli (0, 4.9 ± 5.2, and 4.5 ± 7.8) goats (P < 0.05). As a result, fertilization rates (%) showed a decrease in Angora (50, 100, 24.5, respectively, for cycles 1, 2, and 3) and Honamli (100, 42.5, and 56.3) goats (P < 0.05), whereas recovery rates showed no difference among the different breeds. The methodology presented in this study was found to be an efficient technique for superovulation of the Angora, Kilis, and Honamli goats. Because Hair and Norduz are relatively small breeds, the dosage of FSH might have had a negative effect on the superovulation and embryo yield. Additionally, the use of intra-abdominal washing solutions for preventing adhesions as observed in previous works (data not shown) is believed to have a positive effect on achieving high levels of efficiency in in vivo embryo production.

In vivo embryo recovery vs. in vivo oocyte recovery and in vitro embryo production: Embryo yields and pregnancy rates in cattle

1997 ◽  
Vol 47 (1) ◽  
pp. 364 ◽  
Author(s):  
K.L. Goodhand ◽  
R.G. Watt ◽  
M.E. Staines ◽  
L.C. Higgins ◽  
D.F. Dolman ◽  
...  
Keyword(s):  
Pregnancy Rates ◽  
Embryo Production ◽  
Embryo Recovery ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

331 EFFECT OF SUCESSIVE LAPAROSCOPIC OOCYTE RECOVERY AFTER HORMONAL TREATMENT IN CROSSBRED GOATS

2010 ◽  
Vol 22 (1) ◽  
pp. 321
Author(s):  
S. R. G. Avelar ◽  
K. C. Almeida ◽  
A. F. Pereira ◽  
F. C. Sousa ◽  
R. R. Moura ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Prostaglandin F2α ◽  
Hormonal Treatment ◽  
Ovarian Response ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Hormonal Stimulation ◽  
Exact Test ◽  
Oocyte Recovery

Laparoscopic oocyte recovery (LOR) is a valuable tool for obtaining oocytes for in vitro embryo production. When preceded by a treatment of ovarian stimulation, this technique produces an increase in the amount of oocytes recovered. However, a little information has been found to respect the effect of successive hormonal treatments on both oocyte quantity and quality. Therefore, the objectives of this study were to evaluate the ovarian response and quantitative and qualitative COC production. Five adult crossbred goats were hormonally treated with intravaginal sponges containing 60 mg of medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) for 11 days. In the 8th day of progestagen treatment, 50 μg of prostaglandin F2α analogue (Ciosin, Coopers, São Paulo, Brazil) was administered by i.m. injection. At this time, ovarian stimulation was initiated by the administration of 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five decreasing doses (30/30, 20/20, 20 mg), at 12-h intervals. The animals were fasted for 24 h before the laparoscopic procedure, which was performed just after the sponge removal. A laparoscopic system connected to a 22-gauge needle (WTA, Watanabe, Brazil) and a vacuum pump (Biovacuum, Biocom, Brazil) providing 30 mm Hg was used. All follicles with a size larger than 2 mm present in both ovaries were counted and aspirated. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin (40 μg mL-1). The COCs were graded based on presence of cumulus cells and cytoplasm homogeneity (I to IV). The hormonal treatment and LOR procedures were repeated three times at 14-day intervals. Data were expressed in percentage or mean ± SEM. The differences were analyzed using ANOVA and Tukey’s or Fischer’s exact test when appropriate, with P < 0.05. No statistical differences were found (P > 0.05) for the number of follicles obtained in each LOR session: 17.0 ± 3.91, 18.75 ± 2.59, and 18.0 ± 4.73, respectively. Repeated LOR procedures also did not affect (P > 0.05) the number of aspirated follicle (15.0 ± 3.92, 15.5 ± 2.33, and 16.0 ± 4.36), resulting from the three sessions, respectively. Average recovery rates were not statistically different (P > 0.05), resulting in 74.7%, 62.9%, and 64.6% between sessions. With respect to the percentage of viable COCs (GI and GII) were not observed statistical differences (P > 0.05), as verified the follow values at 1st to 3rd sessions: 76.79%, 84.62%, and 74.19%. In conclusion, three successive hormonal stimulation LOR procedures did not cause detrimental effects on quantitative and qualitative oocyte production, suggesting that this protocol can be used for programs of in vitro goat embryo production. This study was supported the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

scholarly journals Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 216
Author(s):  
Hernan Baldassarre
Keyword(s):  
Pregnancy Rate ◽  
Commercial Application ◽  
Embryo Production ◽  
Breeding Programs ◽  
Sheep And Goats ◽  
Embryo Recovery ◽  
Marker Selection ◽  
Holstein Calves ◽  
In Vitro Embryo Production

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

scholarly journals Prostaglandin F2α Represses IGF-I-Stimulated IRS1/Phosphatidylinositol-3-Kinase/AKT Signaling in the Corpus Luteum: Role of ERK and P70 Ribosomal S6 Kinase

2010 ◽  
Vol 24 (3) ◽  
pp. 632-643 ◽  
Author(s):  
Edward Arvisais ◽  
Xiaoying Hou ◽  
Todd A. Wyatt ◽  
Koumei Shirasuna ◽  
Heinrich Bollwein ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Prostaglandin F2α ◽  
Serine Phosphorylation ◽  
Phosphatidylinositol 3 Kinase ◽  
Akt Activation ◽  
Diminished Capacity ◽  
Igf I ◽  
In Vitro Treatment

Abstract Little is known about the early intracellular events that contribute to corpus luteum regression. Experiments were designed to determine the effects of prostaglandin F2α (PGF2α) on phosphatidylinositol-3-kinase (PI3K)/Akt signaling in the corpus luteum in vivo and in vitro. Treatment of midluteal-phase cows with a luteolytic dose of PGF2α resulted in a rapid increase in ERK and mammalian target of rapamycin (mTOR)/p70 ribosomal protein S6 kinase (p70S6K1) signaling and a rapid suppression of Akt phosphorylation in luteal tissue. In vitro treatment of primary cultures of luteal cells with PGF2α also resulted in an increase in ERK and mTOR/p70S6K1 signaling and a diminished capacity of IGF-I to stimulate PI3K, Akt, and protein kinase C ζ activation. Accounting for the reductions in PI3K and Akt activation observed in response to PGF2α treatment, we found that PGF2α promoted the phosphorylation of serine residues (307, 612, 636) in the insulin receptor substrate 1 (IRS1) peptide sequence in vivo and in vitro. Serine phosphorylation of IRS1 was associated with reduced formation of IGF-I-stimulated IRS1/PI3Kp85 complexes. Furthermore, treatment with inhibitors of the MAPK kinase 1/ERK or mTOR/p70S6K1 signaling pathways prevented PGF2α-induced serine phosphorylation of IRS1 and abrogated the inhibitory actions of PGF2α on Akt activation. Taken together, these experiments provide compelling evidence that PGF2α treatment stimulates IRS1 serine phosphorylation, which may contribute to a diminished capacity to respond to IGF-I. It seems likely that the rapid changes in phosphorylation events are among the early events that mediate PGF2α-induced corpus luteum regression.

scholarly journals Investigating the Intrafollicular Factors Affecting Oocyte Developmental Competency

2021 ◽  
Author(s):  
◽  
Zaramasina Clark
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Significant Finding ◽  
High Quality ◽  
Final Maturation

<p>The number of cycles of assisted reproductive technologies (ART) performed increased by ~9.5 % globally between 2008 and 2010. In spite of this, the success rate in terms of delivery was only ~19.0 % (Dyer et al., 2016). This discrepancy between the demand for, and success of, these technologies necessitates the development of tools to improve ART efficiency. To facilitate this, a better understanding of how the microenvironment changes within the developing follicle to culminate in a mature, developmentally-competent oocyte is required. This study employed an in vivo and in vitro ovine model to investigate the relationship between the surrounding microenvironment and oocyte maturation, and in particular, the attainment of oocyte developmental competency and high-quality embryos.  The first objective of this PhD study was to comprehensively investigate the changing microenvironment of in vivo matured, presumptive preovulatory (PPOV) follicles from wild-type (++) and high ovulation rate (OR; I+B+) ewes. The high OR ewes were heterozygous carriers of mutations in BMP15 (I+) and BMPRIB (B+). Functional differences in follicular somatic (granulosa and cumulus) cells between these genotypes, including differential gonadotropin responsiveness of granulosa cells, composition of follicular fluid and gene expression profiles in cumulus cells were evident. These differences emerged as part of a compensatory mechanism by which oocytes from smaller follicles, containing fewer granulosa cells, achieved developmental competency in I+B+ ewes.  The second objective of this PhD study was to develop new approaches for improving current in vitro maturation (IVM) strategies. The first approach utilised in this study focused on developing biomarkers that could be used to improve prediction of developmental competency in oocytes and in vitro produced embryos. This involved interrogating the hypothesis that a combination of molecular and morphokinetic biomarkers would better predict the developmental competency of oocytes and embryos compared to using these biomarkers alone. The second approach utilised in this PhD study tested the effects of modulating IVM conditions to better mimic the follicular microenvironment of a high, compared to a low, OR species on oocyte developmental competency and embryo quality. This involved supplementing IVM media with different ratios of two oocyte-secreted growth factors, i.e. GDF9:BMP15, that were representative of low or high OR species. These approaches demonstrated significant potential and warrant further investigation.  The most significant finding of this study was that despite variances in the surrounding microenvironment during in vivo and in vitro oocyte maturation that culminated in differential gene expression patterns in cumulus cells, and divergent gonadotropin-responsiveness of granulosa cells, the gene expression signatures of developmentally-competent oocytes and the morphokinetics of high-quality embryos were unaltered. This confirms the value of developing such biomarkers for oocyte development competency and embryo quality that remain unaltered despite a changing surrounding environment. Interestingly, simulating the ratio of GDF9:BMP15 that oocytes from high OR species are exposed to during maturation improved developmental competency in oocytes as demonstrated by increased blastocyst rates. Furthermore, this study has demonstrated that combinations of molecular (cumulus cell gene expression) and morphokinetic biomarkers improved the ability to predict developmental competency in oocytes and embryos. Overall, this study revealed novel information regarding the follicular microenvironment during final maturation and identified several novel approaches to improving the efficiency of ART.</p>

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