scholarly journals Oocytes recovered from cows treated with retinol become unviable as blastocysts produced in vitro

Reproduction ◽  
2005 ◽  
Vol 129 (4) ◽  
pp. 411-421 ◽  
Author(s):  
C Hidalgo ◽  
C Díez ◽  
P Duque ◽  
J M Prendes ◽  
A Rodríguez ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Follicular Fluid ◽  
Teratogenic Effect ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Blastocyst Development ◽  
Time Limits ◽  
Oocyte Donors ◽  
First Session

Retinoids have been shown to enhance developmental competence of the oocyte in cattle, sheep and pigs. In this study we investigated whether exogenous retinol stimulates the bovine oocyte during its intrafollicular growth and the time limits of exposure to exogenous retinol. In addition, we also determined the efficiency of ovum pick-up techniques in combination with retinol treatment and the viability of embryos after transfer to recipients. In Experiment 1, heifers were injected with retinol or vehicle, and concentrations of retinol in the blood were analysed on Day 0 (prior to injection), Day 1 and, together with follicular fluid, Day 4. Blood retinol increased by Day 1 and cleared on Day 4, but retinol remained higher within the follicle. In Experiment 2, oocyte donors were injected weekly with retinol or vehicle four times during a twice-per-week cycle of eight recovery sessions (starting 4 days before the first session), followed by a second eight-session cycle without treatment. Oocytes recovered were fertilized and culturedin vitro.Retinol treatment yielded higher numbers of low-quality oocytes throughout, although retinol measured during cycles did not change. Total oocytes, and morulae and blastocyst rates, increased during the first five sessions following treatment with retinol. As previously shown with oocytes from slaughterhouse ovaries, retinoic acid stimulated blastocyst development. Following transfer to recipients, blastocysts from oocytes exposed to retinol were unable to establish pregnancy. Our study confirms the existence of an effect of retinol on the intrafollicular oocyte in the cow and provides evidence regarding the teratogenic effect of retinol.

207 IN VITRO MATURATION TREATMENT AFFECTS DEVELOPMENTAL COMPETENCE OF LAPAROSCOPIC OVUM PICKUP-DERIVED OOCYTES IN FOLLICLESTIMULATING HORMONE-STIMULATED GOATS

2008 ◽  
Vol 20 (1) ◽  
pp. 182 ◽  
Author(s):  
Y. Locatelli ◽  
N. Poulin ◽  
G. Baril ◽  
J.-L. Touzé ◽  
A. Fatet ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Follicular Fluid ◽  
Developmental Competence ◽  
Postmortem Changes ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Epidermal Growth

The aim of the present study was to assess the effect of IVM treatment on the developmental competence of oocytes recovered from repeated laparoscopic ovum pickukp (LOPU) in goats. A total of 94 LOPU sessions were performed on 33 adult goats of the Saanen and Alpine breeds. Females were synchronized (Day 0) during the nonbreeding season by inserting vaginal sponges (45 mg of fluorogestone acetate, Intervet, Boxmeer, The Netherlands). At Day 8, an i.m. injection of 50 μg of cloprostenol (Estrumate; Schering-Plough Animal Health, Pointe-Claire, Quebec, Canada) was administered. Porcine FSH (Stimufol, Merial, Brussels, Belgium, 160 mg/goat) was administered in 5 injections at 12-h intervals, starting on Day 8. The LOPU took place under general anesthesia on Day 11, and follicles ≥2 mm were aspirated with an 18-gauge needle connected to a controlled vacuum system. Vaginal sponges were removed at the time of LOPU. Treatments were repeated 2 times in a 2-week interval scheme (2 goats and 1 goat were excluded from the experiment during the second and third LOPU sessions, respectively). Cumulus–oocyte complexes were washed and evaluated for quality (graded from 1 to 3). Oocytes recovered from unstimulated slaughterhouse-derived ovaries served as a control. Cumulus–oocytes complexes from Grades 1 and 2 were submitted to IVM in TCM-199, supplemented with 100 μm of cysteamine and either 10 ng mL–1 of epidermal growth factor (EGF) or 10% follicular fluid and 100 ng mL–1 of ovine FSH (FF-FSH). Matured oocytes were then submitted to IVF and in vitro development as described by Cognié et al. (2004 Reprod. Fertil. Dev. 16, 437–445). Over the 94 LOPU sessions, 20.4 ± 0.9 follicles were aspirated (mean ± SEM), allowing the recovery of 12.3 ± 0.7 COC per goat and per session, of which 80.1% were suitable for IVM (Grades 1 and 2). Results of in vitro production are detailed in the table. The IVM treatment did not significantly affect cleavage or blastocyst development rates in oocytes derived from slaughterhouse ovaries. Cleavage rates were significantly decreased in LOPU-derived oocytes when compared with control oocytes. For LOPU-derived oocytes, cleavage and final blastocyst development rates were increased significantly and kinetics of embryo development were accelerated when FF-FSH was used during IVM as compared with EGF. The IVM with FF-FSH allowed us to produce 4.1 blatocysts per goat per LOPU session. These results demonstrate the interest in LOPU for goat embryo production once appropriate IVM treatment is used. The difference observed between LOPU and slaughterhouse oocytes in terms of response to IVM treatments may be related to FSH stimulation prior to the LOPU session or to postmortem changes in oocyte responsiveness in the slaughterhouse group. Table 1. Effects of oocyte origin [laparoscopic ovum pickukp (LOPU) or slaughterhouse derived] and maturation treatment [epidermal growth factor (EGF) or follicular fluid (FF)-FSH] on in vitro embryo production (6 replicates)

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 Pregnancies and improved early embryonic development with bovine oocytes matured in vitro with 9-cis-retinoic acid

Reproduction ◽  
2003 ◽  
pp. 409-416 ◽  
Author(s):  
CO Hidalgo ◽  
C Diez ◽  
P Duque ◽  
N Facal ◽  
E Gomez
Keyword(s):  
Retinoic Acid ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Developmental Potential ◽  
Vitro Fertilization ◽  
Cell Counts ◽  
Bovine Oocytes ◽  
Number Of Cells

Retinoids have an important role in cell growth, morphogenesis and differentiation. In the present study the developmental potential of bovine oocytes was examined after in vitro maturation in the presence of 9-cis-retinoic acid, a vitamin A metabolite, at 5 nmol l(-1) in chemically defined conditions. Experiments studied early in vitro development, blastocyst differential cell counts and the capacity of embryos to establish pregnancy after transfer to recipients. After in vitro fertilization and culture in simple medium, blastocyst development and hatching rates increased in oocytes matured with 9-cis-retinoic acid. Addition of ethanol (used as a solvent for 9-cis-retinoic acid) resulted in higher cell counts and proportions of cells in the inner mass of day 7 blastocysts. Day 8 blastocysts represented most differences observed in the number of cells. In these embryos, ethanol and 9-cis-retinoic acid increased both the number of cells and proportions in the inner mass. However, while ethanol treatment reduced the number of cells in the trophectoderm, 9-cis-retinoic acid did not. The total number of cells was unaffected by treatment within 1 day, although untreated oocytes lead to day 8 blastocysts with reduced total cell counts. Once transferred to recipients, both fresh and vitrified-warmed blastocysts derived from oocytes matured with 9-cis-retinoic acid yielded more pregnancies at day 60. Modifications of retinoid metabolism affect development and trophectoderm differentiation, and in vitro maturation with 9-cis-retinoic acid increased the developmental competence of the oocyte.

scholarly journals 127 DEVELOPMENT AND QUALITY OF BOVINE MORULAE CULTURED IN SERUM-FREE MEDIUM WITH RETINOIC RECEPTOR SPECIFIC AGONISTS

2008 ◽  
Vol 20 (1) ◽  
pp. 144
Author(s):  
E. Gómez ◽  
J. N. Caamaño ◽  
M. Muñoz ◽  
A. Rodríguez ◽  
N. Facal ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Inner Cell Mass ◽  
Retinoic Acid Receptor ◽  
Cell Mass ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Cell Counts ◽  
All Trans Retinoic Acid ◽  
Apoptosis And Necrosis

In the cell, all-trans retinoic acid (ATRA), a vitamin A metabolite, binds to retinoic acid-receptor (RAR), whereas the ATRA isomere 9-cis-retinoic acid (9-cis-RA) binds to both RAR and the retinoid X receptor (RXR). Synthetic compounds such as LG100268 (LG; Ligand Laboratories) are highly specific to bind RXR, which allows to differentially study the RAR and RXR pathways. In previous work morulae treated with LG for 48 h showed to improve blastocyst development and to activate pro-apoptotic genes (in press), whereas ATRA for 24 h increased cell numbers in the inner cell mass (ICM) and the trophectoderm (TE) (Rodr�guez et al. 2006 Hum. Reprod. 21, 2149–2157). However, LG and ATRA were never both compared for 24 in medium with BSA, which is thought to be more appropriate to produce embryos for cryopreservation than serum-containing medium. In this work we analyze development, quality, and viability of morulae cultured with RAR and RXR agonists. Cumulus–oocyte complexes from slaughterhouse ovaries were matured and fertilized in vitro. Presumptive zygotes were cultured in synthetic oviduct fluid (SOF) +3 gL–1 BSA. On day 6, morulae were treated for 24 h with ATRA 0.7 µm, LG 0.1 µm, or no additives. Blastocyst development was monitored up to day 8. Differential cell counts were made on hatched blastocysts on days 7 and 8. Apoptosis and necrosis (TUNEL + nuclear histology) were made on day 8 expanded and hatched blastocysts. Data were analyzed by GLM and Duncan's test, expressed as LSM � SE, and development rates were expressed as percentages of cultured morulae (replicates [R] = 14 for development; R = 9 for cell counts; R = 4 for apoptosis; n = 1647 morulae). ATRA yielded more blastocysts on day 8 than LG and controls (72.2 � 2.2 v. 60.0 � 2.3 and 65.6 � 2.4, respectively; P < 0.02), and more expanded blastocysts than LG (48.6 � 2.3 v. 36.6 � 2.4; P < 0.02), but no more than controls (43.5 � 2.5). Day-7 and day-8 hatched blastocysts cultured with ATRA showed more total cells than day-7 controls (163.5 � 8.0 and 161.5 � 5.4 v. 137.7 � 8.9, respectively; P < 0.05). However, in the presence of ATRA, day-8 blastocysts showed a strong cell reduction in the inner cell mass (ICM), whereas their day-7 counterparts conserved ICM/total cells proportions comparable to day-7 controls (11.0 � 1.2 v. 19.7 � 1.7 and 20.6 � 1.9, respectively; P < 0.03). The LG increased apoptotic index (AI) and necrotic index (NI) in the ICM (AI: 14.5 � 2.4 v. 6.4 � 1.5 and 6.4 � 1.4; NI: 5.0 � 1.2 v. 0.9 � 0.8 and 1.6 � 0.7; for LG, ATRA, and controls, respectively; P < 0.02). Embryos produced with ATRA showed improved development and cell distribution without increasing apoptosis and necrosis. Vitrification of excellent day-7 and day-8 blastocysts is in course to evaluate cryosurvival and further embryo transfer to determine full developmental competence. Grant Support: MEC, project AGL2005-04479. M. Muñoz is sponsored by FICYT.

The effects of brain-derived neurotrophic factor and metformin on in vitro developmental competence of bovine oocytes

Zygote ◽  
2009 ◽  
Vol 17 (3) ◽  
pp. 187-193 ◽  
Author(s):  
So Gun Hong ◽  
Goo Jang ◽  
Hyun Ju Oh ◽  
Ok Jae Koo ◽  
Jung Eun Park ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Embryo Development ◽  
Neurotrophic Factor ◽  
Brain Derived Neurotrophic Factor ◽  
Early Embryo ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Significant Difference

SummaryBrain-derived neurotrophic factor (BDNF) signalling via tyrosine kinase B receptors may play an important role in ovarian development and function. It has been reported that metformin elevates the activity of Tyrosine kinase receptors and may amplify BDNF signalling. The objective of this study was to investigate the effect of BDNF during in vitro maturation (IVM) and/or in vitro culture (IVC) (Experiment 1), and to evaluate the collaborative effect of BDNF and metformin treatment on the developmental competence of bovine in vitro fertilized (IVF) embryos (Experiment 2). In Experiment 1, BDNF, which was added to our previously established IVM systems, significantly increased the proportions of MII oocytes at both 10 ng/ml (86.7%) and 100 ng/ml (85.4%) compared with the control (64.0%). However, there was no statistically significant difference in blastocyst development between the control or BDNF-supplemented groups. In Experiment 2, in order to investigate the effect of BDNF (10 ng/ml) and/or metformin (10−5 M) per se, TCM-199 without serum and hormones was used as the control IVM medium. The BDNF (48.3%) and BDNF plus metformin (56.5%) significantly enhanced the proportions of MII oocytes compared with the control (34.4%). Although, BDNF or metformin alone had no effect in embryo development, BDNF plus metformin significantly improved early embryo development to the 8–16-cell stage compared with the control (16.5 vs. 5.5%). In conclusion, the combination of BDNF and metformin may have a collaborative effect during the IVM period. These results could further contribute to the establishment of a more efficient bovine in vitro embryo production system.

scholarly journals PTPN11 (SHP2) Is Indispensable for Growth Factors and Cytokine Signal Transduction During Bovine Oocyte Maturation and Blastocyst Development

Cells ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 1272 ◽  
Author(s):  
Muhammad Idrees ◽  
Lianguang Xu ◽  
Seok-Hwan Song ◽  
Myeong-Don Joo ◽  
Kyeong-Lim Lee ◽  
...  
Keyword(s):  
Growth Factors ◽  
Embryo Development ◽  
Oocyte Maturation ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Specific Inhibitor ◽  
Mrna Translation ◽  
Bovine Oocyte ◽  
Blastocyst Development

This study was aimed to investigate the role of SHP2 (Src-homology-2-containing phosphotyrosine phosphatase) in intricate signaling networks invoked by bovine oocyte to achieve maturation and blastocyst development. PTPN11 (Protein Tyrosine Phosphatase, non-receptor type 11) encoding protein SHP2, a positive transducer of RTKs (Receptor Tyrosine Kinases) and cytokine receptors, can play a significant role in bovine oocyte maturation and embryo development, but this phenomenon has not yet been explored. Here, we used different growth factors, cytokines, selective activator, and a specific inhibitor of SHP2 to ascertain its role in bovine oocyte developmental stages in vitro. We found that SHP2 became activated by growth factors and cytokines treatment and was highly involved in the activation of oocyte maturation and embryo development pathways. Activation of SHP2 triggered MAPK (mitogen-activated protein kinases) and PI3K/AKT (Phosphoinositide 3-kinase/Protein kinase B) signaling cascades, which is not only important for GVBD (germinal vesical breakdown) induction but also for maternal mRNA translation. Inhibition of phosphatase activity of SHP2 with PHPS1 (Phenylhydrazonopyrazolone sulfonate 1) reduced oocytes maturation as well as bovine blastocyst ICM (inner cell mass) volume. Supplementation of LIF (Leukemia Inhibitory Factor) to embryos showed an unconventional direct relation between p-SHP2 and p-STAT3 (Signal transducer and activator of transcription 3) for blastocyst ICM development. Other than growth factors and cytokines, cisplatin was used to activate SHP2. Cisplatin activated SHP2 modulate growth factors effect and combine treatment significantly enhanced quality and rate of developed blastocysts.

scholarly journals Delaying meiotic resumption during transportation of bovine cumulus–oocyte complexes: effects on development, apoptosis and caspases activity of in vitro-produced embryos

Zygote ◽  
2017 ◽  
Vol 25 (6) ◽  
pp. 740-750 ◽  
Author(s):  
Priscila Chediek Dall'Acqua ◽  
Beatriz Caetano da Silva Leão ◽  
Nathália Alves de Souza Rocha-Frigoni ◽  
Fernanda Patrícia Gottardi ◽  
Gisele Zoccal Mingoti
Keyword(s):  
Bovine Serum Albumin ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
The Other ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Control Groups ◽  
Meiotic Inhibitors

SummaryThis study examined the effects of meiosis inhibition during bovine oocyte transportation on developmental competence and quality of produced embryos. The transportation medium was supplemented with: 100 μM butyrolactone I (BL), 500 μM IBMX + 100 μM forskolin (mSPOM), 100 μM milrinone (MR) or follicular fluid (bFF), and was carried out in a portable incubator for 6 h. Next, oocytes were in vitro matured (IVM) for 18 h, without the meiotic inhibitors, with the exception of mSPOM group, in which was added 20 μM cilostamide. The three control groups were IVM with 10% fetal calf serum (FCS) (Control Lab FCS) or 0.6% bovine serum albumin (BSA) (Control Lab BSA) in a CO2 in air incubator or in the portable incubator with 0.6% BSA (Control Transp BSA). Higher cleavage rates (P < 0.05) were obtained in the Control Lab FCS group (84.5 ± 5.3%) compared with the other groups (59.6 ± 3.4% to 70.9 ± 2.3%). Embryonic development was higher (P < 0.05) in the Control Lab FCS group (39.8 ± 4.7%) than in the Control Transp BSA (22.7 ± 3.4%) and MR (21.6 ± 2.3%) groups. However, they were similar (P > 0.05) to the other groups (23.6 ± 3.3% to 28.8 ± 2.7%). The total number of blastomeres was higher (P < 0.05) in the Control Lab FCS group (85.2 ± 5.6) than in Control Lab BSA (53.6 ± 2.9), Control Transp BSA (55.5 ± 4.4), BL (58.2 ± 3.0), mSPOM (57.9 ± 4.9) and MR (59.2 ± 3.9), but all these treatments did not differ (P > 0.05) from bFF (67.7 ± 4.2). No differences (P > 0.05) were found in apoptosis by the activity of caspases (139.0 ± 3.2 to 152.4 ± 6.5, expressed in fluorescence intensity) as well as the percentage of TUNEL-positive cells (12.3 ± 2.0% to 15.7 ± 1.7%). In conclusion, the transportation of oocytes over 6 h with BL, mSPOM or bFF enabled the acquisition of developmental competence at similar rates to the Control Lab FCS group.

327 OOCYTE-SECRETED FACTORS DIRECTLY AFFECT OOCYTE DEVELOPMENTAL COMPETENCE DURING IN VITRO MATURATION OF THE BOVINE CUMULUS - OOCYTE COMPLEX

2006 ◽  
Vol 18 (2) ◽  
pp. 271 ◽  
Author(s):  
T. S. Hussein ◽  
R. B. Gilchrist ◽  
J. G. Thompson
Keyword(s):  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Paracrine Factors ◽  
Cell Functions ◽  
Oocyte Developmental Competence ◽  
Secreted Factors

Paracrine factors secreted by the oocyte (oocyte-secreted factors, OSFs) regulate a broad range of cumulus cell functions including proliferation, differentiation, and apoptosis. The capacity of oocytes to regulate their own microenvironment by OSFs may in turn contribute to oocyte developmental competence. The aim of this study was to determine if OSFs have a direct influence on bovine oocyte developmental competence during in vitro maturation (IVM). Cumulus-oocyte complexes (COCs) were obtained by aspiration of >3-mm follicles from abattoir-derived ovaries. IVM was conducted in Bovine VitroMat (Cook Australia, Eight Mile Plains, Brisbane, Australia) supplemented with 0.1 IU/mL rhFSH for 24 h under 6% CO2 in air at 38.5�C. In the first experiment, COCs were co-cultured with denuded oocytes (DOs, 5/COC in 10 �L) beginning at either 0 or 9-h of IVM. To generate the 9-h DO group, COCs were first cultured intact for 9-h and then denuded. In the second experiment, specific OSFs, recombinant bone morphogenetic protein-15 (BMP-15) and growth differentiation factor 9 (GDF-9), were prepared as partially purified supernatants of transfected 293H cells, and used as 10% v/v supplements in Bovine VitroMat. Treatments were: (1) control (no supplement), (2) BMP-15, (3) GDF-9, (4) BMP-15 and GDF-9, and (5) untransfected 293H control. Following maturation, in vitro production of embryos was performed using the Bovine Vitro system (Cook Australia) and blastocysts were examined on Day 8 for development. Developmental data were arcsine-transformed and analyzed by ANOVA, followed by Tukey's test. Cell numbers were analyzed by ANOVA. Co-culturing intact COCs with DOs from 0 or 9 h did not affect cleavage rate, but increased (P < 0.001) the proportion of cleaved embryos that reached the blastocyst stage post-insemination (50.6 � 1.9 and 61.3 � 1.9%, respectively), compared to COCs cultured alone (40.7 � 1.4%). Therefore, paracrine factors secreted by DOs increased the developmental competence of oocytes matured as COCs. OSFs also improved embryo quality, as co-culture of COCs with DOs (0 or 9 h) significantly increased total cell (156.1 � 1.3 and 159.1 � 1.3, respectively) and trophectoderm (105.7 � 1.3 and 109.8 � 0.4, respectively) numbers, compared to control COCs (total = 148 � 1.2, trophectoderm = 98.2 � 0.8, P < 0.001). BMP-15 alone or with GDF-9 also significantly (P < 0.001) increased the proportion of oocytes that reached the blastocyst stage post insemination (57.5 � 2.4% and 55.1 � 4.5%, respectively), compared to control (41.0 � 0.9%) and 293H-treated (27.1 � 3.1%) COCs. GDF-9 also increased blastocyst yield (49.5 � 3.9%) but not significantly. These results are the first to demonstrate that OSFs, and particularly BMP-15 and GDF-9, directly affect bovine oocyte developmental competence. These results have far-reaching implications for improving the efficiency of IVM in domestic species and human infertility treatment, and support the role of OSF production by oocytes as a diagnostic marker for developmental competence.

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