Developmental Competence Of Cumulus-oocyte Complexes Research Articles

Oocyte Quality, In Vitro Fertilization and Embryo Development of Alpaca Oocytes Collected by Ultrasound-Guided Follicular Aspiration or from Slaughterhouse Ovaries.

Simple SummaryWe compared the morphological quality and the in vitro developmental competence of cumulus–oocyte complexes (COCs) from live vs. slaughtered alpacas. COCs were recovered using: (i) manual aspiration in slaughterhouse ovaries (n = 15 females), or (ii) ovum pick-up (OPU) in live superstimulated alpacas (n = 13). COCs recovered were morphologically evaluated. Grade I–III COCs were in vitro matured and fertilized afterwards using fresh alpaca spermatozoa. Zygotes were in vitro cultured for seven days. The proportion of COCs recovered was similar between groups, but the mean number of COCs collected from individual ovaries was greater (p < 0.05) in slaughterhouse ovaries. A higher (p < 0.05) percentage of grades III and IV and a lower (p < 0.05) percentage of grade I COCs was obtained using OPU. Although, no significant difference in the percentage of cleavage and morula formation was observed between groups, the number of blastocysts, regarding cleavage and COCs collected, was higher (p < 0.007 and p < 0.0002, respectively) in the OPU group. However, the total number of blastocysts per female did not differ between groups. The recovery rate and morphological quality of COCs was significantly higher when follicles were manually aspirated from slaughterhouse alpaca ovaries; however, a greater developmental potential was observed in oocytes obtained from live alpaca.The morphological quality and the in vitro developmental competence of cumulus-oocyte complexes (COCs) collected from in vivo or slaughtered alpacas was compared. COCs were recovered from ovarian follicles using: (i) manual aspiration in ovaries of alpacas (n = 15) sacrificed at a local slaughterhouse, or (ii) transrectal ultrasound-guided follicular aspiration (or ovum-pick-up, OPU) in live alpacas (n = 13) 4 days after the administration of an ovarian superstimulation protocol (200 UI eCG). COCs recovered from both groups were morphologically evaluated and graded. Grade I to III COCs were in vitro matured for 26 h and in vitro fertilized afterwards for 20 h using fresh alpaca epididymal spermatozoa. Presumptive zygotes from both groups were in vitro cultured for 7 days. The proportion of COCs recovered over the total number of follicles punctured was similar between groups, but the mean number of COCs collected from individual ovaries was greater (p < 0.05) in slaughterhouse ovaries. A significantly higher (p < 0.05) percentage of low-quality COCs (grades III and IV) and a lower (p < 0.05) percentage of grade I COCs was obtained using OPU. The number of blastocysts, regarding cleavage and COCs collected, was higher (p < 0.007 and p < 0.0002 respectively) for COCs collected by OPU; however, the total number of blastocysts per female did not differ between groups. We can conclude that the recovery rate and morphological quality of COCs was significantly higher when follicles were manually aspirated from slaughterhouse alpaca ovaries; however, a statistically higher developmental potential was observed in oocytes collected by OPU from live alpaca donors.

Melatonin enhances in vitro developmental competence of cumulus-oocyte complexes collected by ovum pick-up in prepubertal and adult dairy cattle

Bovine oocytes from prepubertal donors have been used for in vitro embryo production to decrease the generation interval. However, reduced cumulus-oocyte competence, mainly attributed to increased apoptosis, has been observed in oocytes/embryos collected from prepubertal donors. Here, we investigated the effects of the potent antioxidative molecule melatonin on cumulus-oocyte competence and embryo development in prepubertal and adult dairy cattle in vitro. A total of fifteen Holstein Friesian calves, six to ten months old (7.6 ± 1.34 months of age). And fifteen adult cows with one to four calvings (2.3 ± 0.96 calvings) were enrolled as ovum pick up (OPU) donors in this study. Cumulus-oocyte complexes (COCs) were cultured either in the presence or absence of melatonin (0.01 nM). The proportion of cleavage stages, blastocysts, and advanced blastocysts was determined. Embryo quality was assessed via differential staining to determine the total embryonic cells and allocation to the inner cell mass (ICM) and trophectoderm (TE) cells. Melatonin treatment yielded a greater percentage of blastocysts compared to the control group, i.e. oocytes from both adult cows (P = 0.0485; 24.8 ± 3.5% vs. 16.0 ± 3.4%, respectively), and from prepubertal donors (P = 0.0007; Melatonin 23.1 ± 5.1% vs. Control: 11.1 ± 3.5%). Adult cows had significantly (P = 0.0370) greater advanced blastocyst rates than those found in the prepubertal group (13.9%± vs. 7.0±%, respectively). Additionally, the number of ICM, total cells, and the ratios ICM: Total, ICM: TE, respectively, were greater (P < 0.05) after melatonin treatment compared with the control group (39.1 ± 2.8, 98.6 ± 5.7, 0.4 ± 0.01, and 0.7 ± 0.04 vs. 27.3 ± 2.9, 81.2 ± 5.8, 0.34 ± 0.01, and 0.52 ± 0.04, respectively). Blastocysts derived from adult cows had a greater number of TE (P = 0.01) and total embryonic cells (P = 0.0095) compared to the prepubertal donor group (63.5 ± 3.2 and 101.05 ± 4.8 vs. 48.9 ± 4.3 and 78.8 ± 6.5, respectively). Nevertheless, embryonic cell counting in embryos derived from prepubertal COCs equated to that observed from adult donors after melatonin exposure. In conclusion, these results indicate that the presence of melatonin during in vitro maturation improves cumulus-oocyte competence, embryo development, and quality by increasing the allocation of embryonic cells to the ICM compartment and the total number of embryonic cells in both adult and prepubertal bovine donors.

A multilevel analysis identifies the different relationships between amino acids and the competence of oocytes matured individually or in groups

High-protein diets contribute to an increase in urea follicular concentrations associated with decreased fertility. Urea has been shown to interfere with the epidermal growth factor (EGF)/EGFR system, which has been shown to have a beneficial effect during in vitro maturation (IVM) of oocytes. Of note, the number of cumulus-oocyte complexes (COCs) in the maturation medium can change the maturation and the developmental competence of COCs. Therefore, it was hypothesized that, the presence of urea and EGF may have a differential effect on the depletion/appearance of AAs and competence of COCs matured individually (I-IVM system) or in groups (G-IVM system). In the G-IVM system, COCs increased consumption (depletion) of AAs compared with other groups in the presence of high-level urea (40 mg/dl) + EGF (10 ng/ml). In the I-IVM system, the non-cleaved COCs depleted more AAs than the cleaved COCs, in particular in the presence of urea. The combination of urea and EGF increased the depletion of AAs in the G-IVM system. However, the EGF abrogated the urea-induced depletion of AAs by the I-IVM COCs. The use of N-acetyl-l-cysteine as an EGFR inhibitor canceled urea-induced depletion of AAs. This shows the inhibiting effect of urea over the EGF/EGFR system. In the presence of urea + EGF, COCs had a lower degree of developmental competence than control in both I- and G-IVM systems. Arginine had the best predictive power to identify highly competent COCs in the G-IVM system, while glutamine was the best predictor of the cleavage in the I-IVM system. In conclusion, this multi-level study shows that COCs matured individually or in groups may have different association with AAs metabolism. These findings provide new insights into the relationships between AA metabolism and the subsequent developmental competence of COCs.

Expression levels of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells can predict in vitro developmental competence of bovine oocytes.

The efficiency of in vitro embryo production technologies would be improved by the development of suitable non-invasive biomarkers that allow the selection of good quality cumulus-oocyte complexes (COCs). The present study used whole, single oocyte culture to investigate whether the expression levels of follicle-stimulating hormone receptor (FSHR), insulin-like factor 1 receptor (IGF1R) and three steroidogenesis-related enzymes (CYP11al, CYP19al and HSD3β) in cumulus cells reflected the developmental competence of COCs. Cumulus cells were collected from single COCs before maturation culture and relative mRNA levels were assessed using real-time PCR. The analysis indicated that mRNAs for FSHR, IGF1R, CYP11al and HSD3β were present at higher levels in cumulus cells from COCs that failed to form blastocysts compared with cumulus cells from COCs that formed blastocysts. Moreover, FSHR and IGF1R mRNA levels were positively correlated with those of genes for steroidogenesis-related enzymes. In conclusion, poor developmental competence of COCs was related to higher expression of FSHR, IGF1R, CYP11al and HSD3β in cumulus cells, which may indicate the advanced differentiation of cumulus cells into granulosa cells.

107 Follicular wave synchronization and FSH stimulation prior to ovum pickup for invitro embryo production

Invitro embryo production (IVP) has become a reliable alternative for genetic improvement in beef and dairy herds. Maximizing embryo yield and pregnancy per donor are key factors. The aim of this study was to compare ovum pickup (OPU) yields, developmental competence of cumulus-oocyte complexes (COCs), and pregnancy rates from Angus donors in a commercial IVP setting. Donors (&amp;gt;4-year-old pluriparous open dry cows) were handled under the same feeding and environmental conditions. Treatment groups were organised as follows: Group 1: no synchronization (SYNCH; n=5); Group 2: SYNCH with no superstimulation (SOV; n=5); Group 3: SYNCH + SOV (n=5) and OPU 36h after last FSH injection; Group 4: SYNCH + SOV (n=5) and OPU 48h after last FSH injection; and Group 5: SYNCH + SOV (n=5) and OPU 72h after last FSH injection. Follicular waves in groups 2, 3, 4, and 5 were synched by gonadotrophin-releasing hormone (GnRH), prostaglandin F2α (PGF), and controlled internal drug release (CIDR). No pre-synch was used. Injections of FSH (pFSH=180mg, Folltropin) were performed IM twice a day, for three days. A minimum of three replicates were performed for each donor. A Mindray DP30V equipped with a micro-convex transducer 5.0-8.5MHz probe, disposable 20-gauge needle, and a flow rate of 15mL min−1 were used for OPU. All visible follicles (Foll) were punctured and retrieved into a 50-mL 36°C warmed tube with media (phosphate-buffered saline, bovine serum albumin (BSA), and heparin). Viable oocytes were classified according to IETS guidelines. The COCs were matured in 100µL of M199 medium supplemented with ALA-glutamine (0.1mM), Na pyruvate (0.2mM), gentamicin (5µgmL−1), epidermal growth factor (50ngmL−1), oFSH (50ngmL−1), bLH (3μgmL−1), cysteamine (0.1mM), and 10% fetal bovine serum (FBS) for 22 to 24h. Fertilization (Day 0) was carried out using highly fertile sires selected by discontinuous 40%/80% layers (PureSperm) and diluted to a final concentration of 1×106 spermmL−1. Matured oocytes were fertilized in 50µL of modified synthetic oviductal fluid (SOF) media supplemented with fructose (90µgmL−1), penicillamine (3µgmL−1), hypotaurine (11µgmL−1), and heparin (10µgmL−1). After 18h, presumptive zygotes were denuded and cultured under low oxygen tension in 50-µL drops of SOF-BSA for 7 days. On Day 3.5, 2% of FBS was added. On Day 7, fresh transferable (grade 1 and 2, IETS standards) blastocysts were implanted into synchronized recipient cows. Around Day 30, ultrasound diagnosis was performed to determined pregnancy rate (PR). We used ANOVA for comparisons of mean values and X2 test for proportions, α=0.05 (Table 1). In conclusion, synchronization, FSH stimulation, and 48-h coasting before OPU in Angus cows increased the number of collected viable oocytes and embryo development rates. More transferrable embryos and higher rates of PR per OPU were obtained using 36- and 48-h coasting, respectively. Table 1.Follicles (Foll), viable oocytes, cleavage, blastocysts, and pregnancy rates (PR) in Angus cows after ovum pickup (OPU) or invitro embryo production (IVP) SYNCH1 FSH Coasting Foll/OPU Oocytes/OPU Viable oocytes Cleavage,% Embryos,% Embryos/OPU PR,% PR/OPU NO NO NO 10.8a 9.0a 8.9a 63a 25a 2.3a 45a 1.0a YES NO NO 10.1a 8.4a 8.1a 73b 27b 2.2a 51b 1.1a YES YES 36H 16.5b 13.8b 13.6b 70c 29b 4.0b 46a 1.8b YES YES 48H 16.1b 13.4b 13.3b 69c 28b 3.8c 52b 2.0b YES YES 72H 15.6b 13.0b 12.6b 64a 22c 2.8d 55b 1.6c a-dValues with different superscripts in the same column differ (P&amp;lt;0.05). 1SYNCH=synchronized.

Effect of cumulus cells of cumulus-oocyte complexes on in vitro maturation, embryonic developmental and expression pattern of apoptotic genes after in vitro fertilization in water buffalo (Bubalus bubalis)

In the present study, the potential of different grades of cumulus-oocyte complexes (COCs) for in vitro maturation (IVM) and embryonic development was assessed. Further, the association of the expression pattern of anti-apoptotic Mcl-1 and pro-apoptotic Bax genes in embryonic development was analyzed. Abattoir derived oocytes were graded into grade A and B based on surrounding cumulus rings. Out of 1050 ovaries, a total number of 770 and 1360, were of grade A and B COCs, respectively, were aspirated. After IVM, grade A COCs had a significantly higher number of polar bodies (92.04 ± 0.60%) as compared to grade B (85.88 ± 0.46%). On IVF and embryo culture, grade A COCs produced the significantly higher rate of cleavage and blastocyst (90.44 ± 0.71% and 41.55 ± 0.96%) as compared to grade B COCs (79.77 ± 0.76% and 30.44 ± 0.96%). The transcriptional analysis of apoptotic genes expression by Real-time PCR revealed a significantly higher expression of Mcl-1 gene in embryos of grade A as compared to grade B, whereas, the relative expression of Bax gene was down-regulated in grade A than grade B embryos. Thus it was concluded that the pattern of apoptotic genes expression in early-stage embryos can be used as a marker gene to predict the developmental competence of COCs.

Lycium barbarum polysaccharide improves the development of mouse oocytes vitrified at the germinal vesicle stage

The cryopreservation of cumulus oocyte complexes (COCs) is an important process in reproductive medicine. However, this process slows both embryo and offspring development. In this study, the effects of maturation medium supplementation with lycium barbarum polysaccharide (LBP) on the developmental competence of vitrified-warmed mouse COCs and the possible mechanisms facilitating this process were investigated. The addition of LBP to the maturation medium improved the maturation rate and enhanced the developmental competence of vitrified-warmed mouse COCs. This may reduce endoplasmic reticulum stress (ERS), preventing cell death and activating both PI3K/AKT and MAPK3/1. In conclusion, cryopreservation lowers the maturation rate and developmental competence of COCs, which can be ameliorated by the application of LBP after warming during maturation.

C-type natriuretic peptide improved vitrified-warmed mouse cumulus oocyte complexes developmental competence

Cryopreservation of cumulus oocyte complexes (COCs) is important for reproductive medicine. However, the vitrified-warmed COCs have lower maturation rate and subsequent developmental competence compared with fresh COCs. The present study was aimed to evaluate the effects of supplementation of the maturation medium with C-type natriuretic peptide (CNP) on the developmental competence of vitrified-warmed mouse COCs. Addition of CNP to the maturation medium improved the maturation rate and enhanced the developmental competence of vitrified-warmed mouse COCs. The reason may be that vitrified COCs led to a decline in cyclic guanosine monophosphate (cGMP) levels. Furthermore, addition of CNP to the maturation medium elevated cGMP levels of the vitrified-warmed COCs. In conclusion, cryopreservation-associated lower maturation rate and developmental competence of COCs may be ameliorated by CNP during maturation culture after warming.

In vitro fertilization and development of cumulus oocytes complexes collected by ultrasound-guided follicle aspiration in superstimulated llamas

The objective was to evaluate the developmental competence of cumulus-oocyte complexes (COC) collected by follicular aspiration in llamas treated with FSH or eCG. Llamas were assigned randomly to two groups (n = 16 per group) and treated, at the time of ovarian follicular wave emergence, with either: 1) 25 mg of FSH im, twice daily for 4 d; or 2) 1000 IU of eCG as a single i.m. dose. The start of gonadotropin treatment was considered Day 0. Both groups were given 5 mg of Armour Standard LH im on Day 6, and COC were collected by follicle aspiration on Day 7. Expanded COC collected from FSH- (n = 157) and eCG-treated llamas (n = 151) were fertilized in vitro using epididymal sperm, and presumptive zygotes were in vitro cultured in SOF medium for 8 d. The FSH and eCG treatment groups did not differ with respect to: the number of follicles ≥7 mm (16.0 ± 2.7 vs 14.0 ± 1.9, respectively; P = 0.5); the number of COC collected (11.5 ± 1.9 vs 9.7 ± 1.2; P = 0.4); the number of expanded COC (9.8 ± 1.4 vs 9.4 ± 1.2; P = 0.8); or the percentage of presumptive zygotes which developed into 2 to 8 cell stage embryos (65.3 vs 63.1), morulas (46.2 vs 42.5), or blastocysts (23.1 vs 20.5; P > 0.05). In conclusion, FSH and eCG treatments were equally effective for recovery of a high number of expanded COC which were used directly for in vitro fertilization. Furthermore, rate of embryo development was not significantly affected by the gonadotropin treatment used.

Lectin-binding sites in isolated equine cumulus-oocyte complexes: Differential expression of glycosidic residues in complexes recovered with compact or expanded cumulus

Equine cumulus-oocyte complexes (COCs) were analyzed by means of 13 lectins to evaluate their glycoconjugate patterns and to verify differences between COCs recovered with compact (Cp) and expanded (Exp) cumulus. Cumulus cells showed a similar staining pattern in both Cp and Exp COCs with all lectins used, except for a higher reactivity with SNA and GSA II in Cp COCs and SBA in Exp COCs. The zona pellucida (ZP) showed (1) uniform staining with MAL II, RCA 120, and SBA in both Cp and Exp COCs, (2) trilaminar binding pattern with WGA as well as higher Con A reactivity in the outer region of both types of COCs, (3) uniform staining with PNA only in Exp COCs, (4) uniform and trilaminar binding pattern with SNA in Cp and Exp COCs, respectively, and (5) major reactivity with GSA II in Exp COCs. Ooplasm showed similar staining intensity with Con A, HPA, GSA I-B 4, and WGA in both Cp and Exp COCs, with stronger reactivity to GSA II in Exp COCs, whereas SNA, UEA I, and LTA binding sites were present only in Cp COCs. Oocyte cortical granules of both Cp and Exp COCs reacted with Con A and WGA. These results suggest that, in the mare, viable (Cp) and atretic (Exp) COCs display different glycoconjugate staining pattern, which may account for the different maturation and developmental competence of COCs.

315 USE OF PERIFOLLICULAR BLOOD FLOW TO PREDICT THE DEVELOPMENTAL COMPETENCE OF BOVINE CUMULUS-OOCYTE COMPLEXES COLLECTED DURING REPEATED OVUM PICKUP SESSIONS ONCE OR TWICE WEEKLY

On average, only 20% of the cumulus–oocyte complexes (COC) develop to the blastocyst stage (Merton et al. 2003 Theriogenology 59, 651–674). An increase in the blood supply to individual follicles appears to be associated with follicular growth rates, whereas a reduction seems to be closely related to follicular atresia (Acosta et al. 2003 Reproduction 125, 759–767). The purpose of this study was to determine whether qualitative perifollicular blood flow changes can be used to predict the developmental competence of COC collected during repeated ovum pickup (OPU) sessions once or twice weekly. Lactating Holstein cows (n = 20) were used as oocyte donors. After dominant follicle removal, OPU was performed twice (group 1, for 3 weeks) or once (group 2, for six weeks) weekly employing a 7.5-MHz transducer (GE 8C-RS) of an ultrasound scanner (GE Logiq Book). Follicle size and Doppler characteristics were recorded by transvaginal ultrasonography just before COC collection using color flow imaging. Owing for technical limitations for measurement of blood flow in small individual follicles, only the presence or absence of blood flow was assessed for each follicle. When a clearly visible blue or red spot (blood flow) was detected in the follicle wall, it was considered as a follicle with detectable blood flow. Follicles with or without detectable blood flow from each individual cow were aspirated separately. After morphological classification of COC, standard protocols for IVP were used for embryo production (Wrenzycki et al. 2001 Biol. Reprod. 65, 323–331). Cleavage and blastocyst rates were recorded at Day 3 and Day 8, respectively. In total, 464 (246 with and 218 without detectable blood flow) and 243 (125 with and 118 without detectable blood flow) follicles ≥3 mm were aspirated in group 1 and group 2, respectively. Morphology of the COC was similar in all groups. Developmental rates for COC stemming from follicles with or without detectable blood flow in group 1 did not show differences for cleavage rates, 54.0% (34/63) and 56.7% (45/81), and for blastocyst rates, 25.4% (16/63) and 22.2% (18/83), respectively. In group 2, the cleavage rates were also similar for COC originating from follicles with and without detectable blood flow, 54.3% (25/46) and 51.5% (34/66). However, developmental rates up to the blastocyst stage did show a significant difference, 23.9% (11/46) and 15.2% (10/66) for COC aspirated from follicles with or without detectable blood flow (P ≤ 0.05). These results show that using COC originating from follicles with detectable perifollicular blood flow collected once weekly may have a higher developmental competence compared to those from follicle without detectable blood flow. Within the detection limits of this study, differences in perifollicular blood flow during repeated OPU sessions once weekly were predictive of oocyte competence. Ruthe Research Farm, Germany, for providing the animals; Masterrind GmbH, Germany, for donation of the semen; and the HW Schaumann Stiftung for financial support.

313 EVALUATION OF COLOR DOPPLER ULTRASONOGRAPHY OF THE PERIFOLLICULAR BLOOD FLOW TO PREDICT THE DEVELOPMENTAL COMPETENCE OF BOVINE CUMULUS - OOCYTE COMPLEXES COLLECTED DURING REPEATED OVUM PICK-UP SESSIONS

The population of cumulus–oocyte complexes (COCs) used in OPU–IVP (ovum pick-up combined with in vitro production of embryos) is largely homogeneous due to repeated sessions resulting in the elimination of dominant and atretic follicles, especially when a 3–4 day interval is used. However, on average only 20% of the COCs develop to the blastocyst stage (Merton et al. 2003 Theriogenology 59, 651–674). Different blood flow changes within the follicle wall influence the fate of the follicles, and detectable blood flow and vasculature are associated with follicle viability. Furthermore, blood flow in follicles may be involved in not only selection of the dominant follicle but also early follicular development including follicular recruitment (Miyamoto et al. 2006 J. Reprod. Dev. 52, 153–160). However, no data are available regarding the quality of the COCs collected from follicles with or without blood flow. The purpose of this study was to determine whether qualitative perifollicular blood flow changes can be used to predict the developmental competence of COCs collected during repeated OPU sessions. Lactating Holstein cows were used as oocyte donors. After dominant follicle removal, OPU was performed twice weekly employing a 7.5-MHz transducer (GE 8C-RS) of an ultrasound scanner (GE Logiq Book). Follicle size and Doppler characteristics (color flow imaging) were recorded by transvaginal ultrasonography just before COC collection. Due to technical limitations for measurement of blood flow in small individual follicles, only the presence or absence of blood flow was assessed for each follicle. When a clearly visible blue or red spot (blood flow) was detected in the follicle wall, it was considered as a follicle with detectable blood flow. Follicles with or without detectable blood flow from each individual cow were aspirated separately. After morphological classification of COCs, standard protocols for IVP were used for embryo production. Cleavage and blastocyst rates were recorded at Day 3 and Day 8, respectively. In total, 464 (246 with and 218 without detectable blood flow) follicles e3 mm were aspirated. The percentage of follicles with detectable blood flow increased depending on follicle size (3 mm: 28.7, 4 mm: 48.4, 5 mm: 50.5, 6 mm: 62.5, 7 mm: 64.8, 8 mm: 71.4, and 9 mm: 76.9). Cleavage rates for COCs stemming from follicles with or without detectable blood flow did not show differences, 45.5% (35/77) and 56.7% (38/67), respectively. The rates of blastocyst formation were also similar in COCs originating from follicles with and without detectable blood flow, 16.9% (13/77) and 14.9% (10/67), respectively. These results show that perifollicular blood flow increases during early follicular growth. Within the detection limits of this study, differences in perifollicular blood flow during repeated OPU sessions twice weekly did not seem to be predictive of oocyte competence. We acknowledge the Ruthe Research Farm, Germany, for providing the animals, and Masterrind GmbH, Germany, for donating the semen.

Effect of ovarian phase and follicle quality on morphology and developmental capacity of the bovine cumulus-oocyte complex.

In order to study the effect of the follicular environment on the quality and developmental competence of cumulus-oocyte complexes (COC), COC were collected from nonatretic (NA), light atretic (LA), atretic (A), and heavily atretic (HA) follicles. Cumulus-oocyte complexes were also collected from early-luteal phase ovaries (EL), from late-luteal phase ovaries (LL), from follicular phase ovaries (F), and from ovaries from non-cyclic animals (NC). Each COC was assigned to one of three quality groups based on the appearance of the cumulus investment: 1) COC-A: compact and bright, 2) COC-B: less compact and dark, and 3) COC-C: strongly expanded cumulus with dark spots. There was a high correlation between follicle quality and the distribution of the COC over the three COC qualities. The COC-A were mainly but not exclusively derived from NA follicles, COC-B were mainly derived from all classes of atretic follicles, and COC-C nearly exclusively originated from HA follicles. The developmental capacity of COC-A and COC-B, which was measured by in vitro embryo production, was consistent over the follicle qualities, except for COC-B obtained from HA follicles. They showed lower development (10.6%, P < .05) compared with COC-B from the other follicle qualities (18.9, 18.7, and 19.8%, respectively, for COC-B from NA, A, and LA follicles). The COC-B from atretic follicles produced more blastocysts (19.8%) than COC-A (12.7%, P < .05). The overall percentage of produced embryos per follicle class seemed to increase with increasing signs of atresia, except if the COC were derived from HA follicles. This increased percentage of embryos was, however, not due to a better quality of COC, but to a higher percentage of COC-B coming from these follicles. The stage of the cycle had no effect on the distribution of the COC over the three COC qualities or on the developmental capacity of COC-A or COC-B, except for COC-A from EL ovaries, which produced more (P < .05) blastocysts than COC-A from the other luteal phases (12.5% vs approximately 8%). A follow-up study was performed trying to elucidate why COC-B possess a higher developmental potency than COC-A. The answer was sought in the oocyte maturation. At several time points during maturation, oocytes were evaluated for their nuclear stage. At all time points COC-B seemed to be a few hours ahead of COC-A, and after 24 h of maturation more COC-B had reached the metaphase-2 stage. This might mean that COC-A need a longer maturation period than COC-B.