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 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.

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)

PSX-A-10 Late-Breaking: Effects of paternal high energy diets on blastocyst development during in vitro embryo production in the bovine

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 363-363
Author(s):  
Dylan B Davis ◽  
Zachary Seekford ◽  
Mackenzie Dickson ◽  
Lucas Gonçalves ◽  
Samir Burato ◽  
...  
Keyword(s):  
High Gain ◽  
High Energy ◽  
Carcass Composition ◽  
Adaptation Period ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Ad Libitum ◽  
In Vitro Embryo Production ◽  
To Receive

Abstract The objective of this study was to evaluate the effect of paternal high energy diets on blastocyst development during in vitro embryo production (IVP). Eight sires were stratified by body weight (initial BW = 946 ± 85 kg) and randomly assigned to the same diet (NEm = 2.10, NEg = 1.44, CP = 14.1%, NDF = 16.6%, DM basis) fed at two different inclusion rates while having ad libitum access to bermudagrass hay (NEm = 1.02, NEg = 0.45, CP = 10.2%, NDF = 71.6). After a 10-d adaptation period, sires were individually fed to receive 0.5% (MAINT) or 1.25% [High gain (HG)] of their BW daily for 67 days. At the end of the feeding period, semen was collected through electroejaculation and frozen. Antral follicles were aspirated from ovaries obtained from a slaughterhouse and utilized for IVP in 4 independent replicates (n = 2,227 total oocytes). Cleavage rates were evaluated 48 h after fertilization and blastocyst development rates were evaluated after 7 days of embryo culture. The proposed treatments successfully induced differences in BW gain (P < 0.01; 2.28 vs -0.04 kg/d) and carcass composition (Rump fat: 1.63 vs. 0.41 cm, P = 0.08; Rib fat: 1.06 vs. 0.41 cm, P = 0.02; intramuscular fat: 3.5 vs. 3.0%, P = 0.36; for HG vs. MAINT sires, respectively). There was a significant decrease in cleavage rates (69.9 ± 2.5 vs. 65.0 ± 2.7; P < 0.04), blastocyst rate as a percentage of oocytes (16.7 ± 2.9 vs. 11.5 ± 2.1; P < 0.01), and blastocyst rates as a percentage of cleaved structures (24.1 ± 3.8 vs. 11.5 ± 2.1; P < 0.01) for HG compared with MAINT sires. In conclusion, sires fed diets that induce highly anabolic conditions had impaired blastocyst development compared to sires fed a maintenance diet.

scholarly journals Oocyte Competence Biomarkers Associated With Oocyte Maturation: A Review

2021 ◽  
Vol 9 ◽  
Author(s):  
Batara Sirait ◽  
Budi Wiweko ◽  
Ahmad Aulia Jusuf ◽  
Dein Iftitah ◽  
R. Muharam
Keyword(s):  
Oocyte Maturation ◽  
Blastocyst Stage ◽  
Current Approach ◽  
Developmental Competence ◽  
Matrix Remodeling ◽  
Oocyte Competence ◽  
Nuclear Maturation ◽  
Oocyte Developmental Competence ◽  
Cumulus Oocyte Complex

Oocyte developmental competence is one of the determining factors that influence the outcomes of an IVF cycle regarding the ability of a female gamete to reach maturation, be fertilized, and uphold an embryonic development up until the blastocyst stage. The current approach of assessing the competency of an oocyte is confined to an ambiguous and subjective oocyte morphological evaluation. Over the years, a myriad of biomarkers in the cumulus-oocyte-complex has been identified that could potentially function as molecular predictors for IVF program prognosis. This review aims to describe the predictive significance of several cumulus-oocyte complex (COC) biomarkers in evaluating oocyte developmental competence. A total of eight acclaimed cumulus biomarkers are examined in the study. RT-PCR and microarray analysis were extensively used to assess the significance of these biomarkers in foreseeing oocyte developmental competence. Notably, these biomarkers regulate vital processes associated with oocyte maturation and were found to be differentially expressed in COC encapsulating oocytes of different maturity. The biomarkers were reviewed according to the respective oocyte maturation events namely: nuclear maturation, apoptosis, and extracellular matrix remodeling, and steroid metabolism. Although substantial in vitro evidence was presented to justify the potential use of cumulus biomarkers in predicting oocyte competency and IVF outcomes, the feasibility of assessing these biomarkers as an add-on prognostic procedure in IVF is still restricted due to study challenges.

246 IN VITRO PRODUCTION OF SPRINGBOK (ANTIDORCAS MARSUPIALIS) EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 239 ◽  
Author(s):  
R. Krisher ◽  
A. Auer ◽  
K. Clark ◽  
K. Emsweller ◽  
S. Rogers ◽  
...  
Keyword(s):  
South Africa ◽  
Oocyte Maturation ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Genetic Management ◽  
Blastocyst Development ◽  
Antidorcas Marsupialis

The objective of this experiment was to develop in vitro embryo production (IVP) technologies in springbok (Antidorcas marsupialis), a southern African antelope. Springbok, a fairly common species on game farms in parts of South Africa, may be used as a model species for gamete rescue and IVP techniques to be applied to the conservation of other threatened antelope species. Springbok belong to the family bovidae, subfamily antilopinae, tribe antilopini, which comprises about twenty species in genera Gazella, Antilope, Procapra, Antidorcas, Litocranius, and Ammodorcas. In this tribe alone, there are 4 species or subspecies that are critically endangered, 3 that are endangered, and 10 that are considered vulnerable, demonstrating the need for antelope conservation efforts. In addition, our studies contributed to the South African biological resource bank, so that banked springbok semen and embryos might be used in the future for managed genetic contribution to isolated captive or wild populations via assisted reproductive technologies. Oocytes were recovered (3 replicates) from ovaries obtained at supervised culls for management purposes in South Africa, and cultured in defined Gmat or undefined TCM-199 with FCS maturation medium for 28-30 h (Brad et al. 2004 Reprod. Fertil. Dev. 16, 223). Oocytes were fertilized with frozen-thawed springbok epididymal spermatozoa in modified SOF fertilization medium with caffeine (Herrick et al. 2004 Biol. Reprod. 71, 948–958). Eighteen hours after insemination, a randomly selected subset of the zygotes were fixed to determine fertilization success. The remaining zygotes were cultured in G1/G2 media. On Day 7 of culture, embryos were analyzed for development to the morula or blastocyst stage. A total of 259 selected oocytes were collected from 50 females (5.2 selected oocytes/female on average). There was no difference in the percentage of oocytes normally fertilized (2 pronuclei, PN) between oocytes matured in Gmat (n= 43; 12%) and those matured in TCM-199 (n= 42; 10%). There were significantly (P < 0.05) more oocytes penetrated (e2 PN) when matured in TCM (50%) compared to Gmat (23%). There were no differences in embryonic cleavage or morula/blastocyst development (of total oocytes inseminated) between treatments (Gmat,n= 89, 54%, 9.0%; TCM-199, n= 85, 68%, 9.4%, respectively). In both treatments, the average blastocyst grade was 2.125 using the standard bovine grading system (Curtis, Cattle Embryo Transfer Procedure, 1991). In conclusion, in vitro oocyte maturation, fertilization, and embryo culture to the blastocyst stage is possible in springbok. Importantly, blastocysts can be produced in vitro under semi-defined conditions, demonstrating that oocyte maturation without serum does support developmental competence. This is important for the potential international movement of IVP embryos to be used for genetic management in the conservation of antelope species.

231 EFFECTS OF DIETARY PROPYLENE GLYCOL ON FOLLICULAR FLUID AND OVUM PICK UP IN VITRO EMBRYO PRODUCTION IN GROWTH-RESTRICTED HEIFERS WITH TWO PROFILES OF ANTI-MÜLLERIAN HORMONE

2015 ◽  
Vol 27 (1) ◽  
pp. 205
Author(s):  
G. Gamarra ◽  
C. Ponsart ◽  
S. Lacaze ◽  
B. Le Guienne ◽  
P. Humblot ◽  
...  
Keyword(s):  
Propylene Glycol ◽  
Follicular Fluid ◽  
Plasma Concentrations ◽  
Oestrous Cycle ◽  
Water Control ◽  
Embryo Production ◽  
Blood Samples ◽  
Metabolic Hormones ◽  
In Vitro Embryo Production

Fertility and embryo quality can be improved in cattle by using diets that induce a programmed modulation of circulating insulin concentrations. The aim of this study was to test whether the daily oral administration of propylene glycol (PG) could modify metabolite and hormone plasma and follicular fluid concentrations and improve in vitro embryo production in superovulated growth-restricted heifers (600 g day–1). Sixteen Holstein heifers were grouped according to their pre-experimental anti-Müllerian hormone (AMH) plasma concentrations: low (L = 1–80 pg mL–1; n = 7) or high (H: >150 pg mL–1; n = 9). Heifers received a single daily drench from Day 1 to Day 9 of an oestrous cycle [first cycle, 400 mL of water (control) and second cycle, 400 mL of PG]. Serial jugular blood samples were collected on Day 7 of each cycle to monitor plasma insulin, glucose, and β-hydroxybutyrate (BHB) concentrations in relation to the drench. Blood samples were also collected to measure insulin-like growth factor-1 (IGF1) and progesterone (P4) concentrations on Days 0, 2, 5, 7, and 9 of the oestrous cycle. Follicular fluid was collected on Day 9 to measure insulin and IGF1 concentrations. Ovarian ultrasonography was performed on Days 2 and 5 to count follicles between 2 and 8 mm in diameter and estimate their size. After ovum pickup (OPU) performed following superovulation on Day 5 of the oestrous cycle, oocytes were matured and fertilized in vitro, then embryos were cultured for 7 days. Propylene glycol increased plasma concentrations of insulin and glucose and reduced BHB in both groups of heifers compared with control. It also increased IGF1 concentrations on Days 5 and 7 in AMH L heifers and on Days 2, 5, and 7 in AMH H heifers, and reduced P4 concentrations on Days 5 and 9 of the oestrous cycle in all heifers. In follicular fluid, there was no difference in insulin concentrations between groups, but PG increased IGF1 concentrations in all heifers. In ovaries, PG increased the number of small follicles (2–3 mm) and total follicles on Day 2 of the cycle in all heifers, and medium follicles (4–8 mm) and total follicles on Day 5 in AMH H heifers. Propylene glycol improved the in vitro embryo development rate (total number of embryos/number of fertilized oocytes) in all heifers (AMH L: control, 37.9% v. PG, 50.0%; P < 0.05; AMH H: control, 36.4% v. PG, 48.3%; P < 0.05). In AMH H, the number of grade 1 blastocysts was increased by PG (control, 5.2 ± 1.0 v. PG, 8.9 ± 1.0; P < 0.01), whereas there was no difference between treatments in AMH L heifers (control, 1.9 ± 1.1 v. PG, 3.2 ± 1.1; P > 0.05). These results indicate that short-term oral PG supplementation affects the concentrations of metabolites and metabolic hormones in blood and IGF1 concentrations in follicular fluid. PG administration is effective in improving in vitro embryo production more markedly in heifers with high AMH compared with low AMH endocrine levels.

342 DISTINCT EFFECTS OF FOLLICULAR FLUID ON CUMULUS CELL APOPTOSIS AND PORCINE OOCYTE DEVELOPMENTAL COMPETENCE

2007 ◽  
Vol 19 (1) ◽  
pp. 286
Author(s):  
C. G. Grupen ◽  
T. S. Hussein ◽  
S. J. Schulz ◽  
D. T. Armstrong
Keyword(s):  
Oxidative Stress ◽  
Follicular Fluid ◽  
Cell Apoptosis ◽  
Polar Body ◽  
Cumulus Cell ◽  
Developmental Competence ◽  
Potent Inducer ◽  
Oocyte Developmental Competence ◽  
Cytoplasmic Maturation

Supplementing medium with follicular fluid (FF) during in vitro maturation (IVM) enhances the developmental competence of porcine oocytes, indicating that factors present in FF are beneficial to cytoplasmic maturation. Previous findings suggest that porcine FF contains high levels of superoxide dismutase activity and exerts a beneficial effect on cytoplasmic maturation by protecting oocytes from oxidative stress (Tatemoto et al. 2004 Biol. Reprod. 71, 1150–1157). Since oxidative stress is a potent inducer of apoptosis, the aim of the present study was to examine the temporal effects of FF during IVM on cumulus cell apoptosis and oocyte developmental competence. Ovaries of prepubertal pigs were collected from a local abattoir and antral follicles, 3 to 7 mm in diameter, were aspirated. Cumulus–oocyte complexes (COCs) with at least 3 uniform layers of compact cumulus cells (CCs) were recovered, washed, and transferred to maturation medium (MM) with or without 25% FF. At 22 h of IVM, COCs from each group were washed and transferred to fresh MM with or without 25% FF, forming 4 groups: -FF/-FF, -FF/+FF, +FF/-FF, and +FF/+FF. Cohorts of COCs were TUNEL stained at 22 and 44 h of IVM using the In Situ Cell Death Detection kit (Roche Diagnostics, Castle Hill, NSW, Australia) according to the manufacturer&apos;s instructions, and apoptotic CCs were visualized using confocal microscopy. Oocytes denuded at 44 h, that had a polar body, were treated with ionomycin and 6-dimethylaminopurine to induce parthenogenetic development, and were cultured for 7 days in NCSU-23 medium at 38.5&deg;C in 5&percnt; O2, 5&percnt; CO2, and 90&percnt; N2. Data were subjected to ANOVA and Tukey&apos;s post-hoc test. At 22 h of IVM, the presence of FF reduced the proportion of apoptotic CCs in COCs (2.1&percnt; vs. 4.6&percnt;). COCs matured with FF from 22 to 44 h of IVM had much lower proportions of apoptotic CCs (&plus;FF/&plus;FF: 0.9&percnt;; &minus;FF/&plus;FF: 2.6&percnt;) compared with those matured without FF (&plus;FF/&minus;FF: 10.3&percnt;; &minus;FF/&minus;FF: 17.8&percnt;). The rate of maturation to the metaphase-II stage was greater when oocytes were matured with FF from 0 to 22 h of IVM (&minus;FF/&minus;FF: 68.6&percnt;; &minus;FF/&plus;FF: 72.8&percnt;; &plus;FF/&minus;FF: 89.2&percnt;; &plus;FF/&plus;FF: 86.2&percnt;). Maturation without FF for the entire IVM interval reduced the proportion of activated oocytes that formed blastocysts compared with the other groups (&minus;FF/&minus;FF: 25.1&percnt;; &minus;FF/&plus;FF: 44.6&percnt;; &plus;FF/&minus;FF: 46.6&percnt;; &plus;FF/&plus;FF: 47.3&percnt;). Despite a 4-fold difference in the proportion of apoptotic CCs between COCs of the &plus;FF/&minus;FF and &minus;FF/&plus;FF groups, exposure to FF for the first or second half of IVM was as beneficial to oocyte developmental competence as exposure to FF for the entire IVM interval. This suggests that the protective effect of FF in reducing oxidative stress on oocytes during IVM is distinct from the effect on oocyte developmental competence.

scholarly journals Endocrine-disrupting chemicals in human follicular fluid impair in vitro oocyte developmental competence

2012 ◽  
Vol 27 (4) ◽  
pp. 1025-1033 ◽  
Author(s):  
Evi M.L. Petro ◽  
Jo L.M.R. Leroy ◽  
Adrian Covaci ◽  
Erik Fransen ◽  
Diane De Neubourg ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Endocrine Disrupting Chemicals ◽  
Developmental Competence ◽  
Human Follicular Fluid ◽  
Oocyte Developmental Competence ◽  
Endocrine Disrupting
Sign in / Sign up

Export Citation Format

Share Document