Overcoming poor in vitro nuclear maturation and developmental competence of domestic cat oocytes during the non-breeding season

The domestic cat experiences circannual variations in ovarian activity and intrafollicular oocyte quality. One result is poor nuclear and cytoplasmic maturation during in vitro maturation (IVM) conducted during the annual non-breeding season (July through November). In an attempt to overcome this seasonal phenomenon immature oocytes were collected from July through November and cultured in a conventional IVM medium (IVM1) or in IVM1 supplemented with different FSH concentrations and antioxidant (ascorbic acid or cysteine). Nuclear status of oocytes was assessed after IVM or IVF. Embryo stage and blastocyst quality were evaluated after 7 days of in vitro culture. Although the addition of antioxidant alone had no effect, the presence of 10 microg FSH ml(-1) improved nuclear maturation (75.4+/-4.1% versus 48.7+/-8.8% in IVM1; P<0.05) and fertilization success (47.9+/-3.2% versus 35.0+/-5.1% in IVM1; P<0.05). Furthermore, developmental competence of fertilized oocytes was enhanced (P<0.05) only in the presence of ascorbic acid (30.6+/-6.7%) or cysteine (33.6+/-5.1%) compared with IVM1 (8.1+/-8.8%). Consequently, blastocyst yield (17% of total oocytes cultured) was highest when oocytes were matured in medium containing higher FSH concentration and antioxidants. The results of this study demonstrate that meiotic and developmental competences are inherent to the immature cat oocyte collected during the non-breeding season. However, appropriate mechanisms (perhaps seasonal variation in FSH receptors or lack of antioxidant capacity of the cumulus-oocyte complex) are inadequate during this period of gonadal quiescence. Regardless, this compromised oocyte function during the non-breeding season can be overridden by altering in vitro culture conditions to include supplemental FSH and antioxidant.

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Overcoming poor in vitro nuclear maturation and developmental competence of domestic cat oocytes during the non-breeding season

Reproduction ◽

10.1530/reprod/126.6.809 ◽

2003 ◽

Vol 126 (6) ◽

pp. 809-816

Author(s):

P Comizzoli

Keyword(s):

Breeding Season ◽

Domestic Cat ◽

Developmental Competence ◽

Nuclear Maturation

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In vitro Culture of Early Secondary Preantral Follicles to Obtain MII Oocyte Developmental Competence from CD-1 Outbred Mice

Indian Journal of Animal Research ◽

10.18805/ijar.b-1247 ◽

2020 ◽

Author(s):

Jongwon Kim ◽

Seungki Lee ◽

Jung Kyu Choi

Keyword(s):

In Vitro Culture ◽

Culture System ◽

Ovarian Follicle ◽

Oocyte Quality ◽

Developmental Competence ◽

Vitro Fertilization ◽

Preantral Follicles ◽

Successful Isolation ◽

In Vitro Culture System

Background: The ovarian follicle is the fundamental functional tissue unit that consists of mammalian ovary. In humans, it has been known that females are born with a maximum number of follicles or oocytes that are not only non-renewable, but also undergoing degeneration with time with a sharply decreased oocyte quality after the age of 35. Methods: Here, we demonstrate that successful isolation of primary, early secondary and late secondary follicles from the ovaries of CD-1 outbred female mice and in vitro culture system to successfully induce the development of MII oocytes. Result: The 9 days of in vitro culture of early secondary follicles showed significant higher rates in growth and maturation displaying higher numbers of antral follicles and MII oocytes developed from early secondary follicles compared to those cultured for 11 days. However, there was no visible difference induced by the size of initial follicles in the rates of growth and maturation. MII oocytes derived from in vitro culture of early secondary follicles following in vitro fertilization developed into two-cell embryos. These observations demonstrate that developmentally competent MII oocytes can be obtained by in vitro culture of preantral follicles derived from the ovaries of CD-1 mice and reveal a crucial role for CD-1 mice as a novel model for research on human ovarian follicles. Furthermore, this study proposes an in vitro culture system using preantral follicle as a therapeutic strategy for fertility preservation of humans for assisted reproductive medicine.

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Early germinal vesicle breakdown is a predictor of high preimplantation developmental competent oocytes in mice

Zygote ◽

10.1017/s0967199416000290 ◽

2016 ◽

Vol 25 (1) ◽

pp. 41-48 ◽

Cited By ~ 2

Author(s):

Shogo Higaki ◽

Masao Kishi ◽

Keisuke Koyama ◽

Masashi Nagano ◽

Seiji Katagiri ◽

...

Keyword(s):

Time Course ◽

Evaluation Method ◽

Germinal Vesicle ◽

Oocyte Quality ◽

Developmental Competence ◽

Germinal Vesicle Breakdown ◽

Nuclear Maturation ◽

Non Invasive ◽

Germinal Vesicle Break Down

SummaryThe preselection of highly developmentally competent oocytes for in vitro maturation (IVM) is crucial for improving assisted reproductive technology. Although several intrinsic markers of oocyte quality are known to be closely related to the onset of nuclear maturation (germinal vesicle break down, GVBD), a direct comparison between GVBD timing and oocyte quality has never been reported. In this study, we established a non-invasive oocyte evaluation method based on GVBD timing for preselecting more developmental competent oocytes in mice. Because the O2 concentration during IVM may affect the nuclear kinetics, all experiments were performed under two distinct O2 concentrations: 20% and 5% O2. First, we determined the time course of changes in nuclear maturation and preimplantation developmental competence of in vitro-matured oocytes to estimate GVBD timing in high developmental competent oocytes. Two-thirds of oocytes that underwent GVBD in early IVM seemed to mainly contribute to the blastocyst yield. To confirm this result, we compared the preimplantation developmental competence of the early and late GVBD oocytes. Cleavage and blastocyst formation rates of early GVBD oocytes (80.2% and 52.7% under 20% O2, respectively, and 67.6% and 47.3% under 5% O2, respectively) were almost double those of late GVBD oocytes (44.8% and 26.0% under 20% O2, respectively, and 40.4% and 17.9% under 5% O2, respectively). With no observable alterations by checking the timing of GVBD in preimplantation developmental competence, oocyte evaluation based on GVBD timing can be used as an efficient and non-invasive preselection method for high developmental competent oocytes.

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hCG Priming Before Ovary Collection Increasing The Oocyte Quality In The Domestic Cat

Acta Veterinaria Indonesiana ◽

10.29244/avi...123-126 ◽

2021 ◽

pp. 123-126

Author(s):

Karisma Mardatillah ◽

Rini Widyastuti ◽

Diah Nugrahani Pristihadi ◽

Wahyudin ◽

Sigit Prastowo ◽

...

Keyword(s):

Oocyte Quality ◽

Domestic Cat ◽

Developmental Competence ◽

Control Group ◽

Oocyte Competence ◽

Slicing Method ◽

Determining Factor

Oocyte competence is a determining factor that influences the embryo development. Embryos produced in vitro have a reduced developmental competence than embryos produced in vivo. Therefore, human Chorionic Gonadotropin (hCG) injection was carried out to improve the quality of the oocytes. The objective of this study was to evaluate the effect of ovarian stimulation with hCG before ovary collection on oocyte quality in the domestic cat. Oocyte donors were either 1) treated with a single dose of 200 IU hCG four days before ovary collection (hCG group), or, 2) no treatment before ovary collection (control group). The oocytes were collected by the slicing method. Immature cumulus oophorus complexes (COCs) from both groups were pooled and matured in vitro for 24-26 hours. Then mature oocytes were fertilized with epididymal sperm and cultured in vitro for seven days. The results study showed that the number of the dominant follicle (DF) and the number of COCs in the hCG group was higher than the control group in right and left ovaries (p<0.05). The morulae and blastocyst rates from cleavage embryos were 88% and 75%, respectively. These results demonstrate that hCG priming of oocytes donors before ovary collection improve oocyte quality.

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26 Season affects cryotolerance of in vitro-produced buffalo embryos

Reproduction Fertility and Development ◽

10.1071/rdv31n1ab26 ◽

2019 ◽

Vol 31 (1) ◽

pp. 139 ◽

Cited By ~ 1

Author(s):

M. A. Kosior ◽

E. Parente ◽

F. Salerno ◽

K. Annes ◽

R. Annunziata ◽

...

Keyword(s):

Developmental Stage ◽

Breeding Season ◽

Embryo Quality ◽

Sucrose Solution ◽

Oocyte Quality ◽

Developmental Competence ◽

Seasonal Effect ◽

Morphological Criteria ◽

Breeding Seasons

Buffaloes are tendentially short-day breeders, and seasonality is one of the main factors affecting the feasibility of ovum pickup and in vitro embryo production technology in this species. An improvement of oocyte developmental competence during decreasing daylight months was previously reported in Italian Mediterranean buffalo (Di Francesco et al. 2011 Anim. Reprod. Sci. 123, 48-53). The aim of this work was to evaluate whether season also affects embryo quality and cryotolerance. Abattoir-derived buffalo cumulus-oocyte complexes were collected during the breeding season, characterised by decreasing daylight length (n=349 over 6 replicates), and the non-breeding season, characterised by increasing daylight length (n=770 over 12 replicates). Buffalo cumulus-oocyte complexes were in vitro matured, fertilized, and cultured according to standard procedures (Di Francesco et al. 2011 Anim. Reprod. Sci. 123, 48-53). The embryos obtained by the end of culture (i.e. on Day 7 post-IVF) were scored for quality and developmental stage, and the percentages of total transferable embryos (tight morulae and blastocysts) were recorded. Embryos (n=107 and 110 in the breeding and non-breeding seasons, respectively) were vitrified by cryotop in 16.5% ethylene glycol, 16.5% dimethyl sulfoxide, and 0.5M sucrose (Boccia et al. 2013 Ital. J. Anim. Sci. 12, 492-496). Warming was carried out by plunging the cryotop strip into a 0.25M sucrose solution and transferring the embryos into 0.15M sucrose for 5min. Embryos were then washed and cultured in SOF for 24h to evaluate post-culture viability. The resistance to cryopreservation was evaluated by assessing the survival rate, on the basis of morphological criteria, and development rate (i.e. the percentage of embryos that resumed their development and reached a more advanced developmental stage) after 24h post-warming culture. Data were analysed by Student’s t-test. Both cleavage (82.8±4.3v. 73.1±1.7 in the breeding and non-breeding seasons, respectively; P<0.05) and blastocyst (32.9±3.5v. 18.3±1.7 in the breeding and non-breeding seasons, respectively; P<0.01) rates increased during the breeding season, confirming previous observations. Due to the different efficiency, a higher number of replicates was required during the non-breeding season to obtain an equal number of embryos. In addition, a seasonal effect was recorded on embryo quality, indicated by poorer cryotolerance of in vitro-produced buffalo embryos during the non-breeding season. Indeed, both survival (94.6±2.7% and 74.0±5.5% in the breeding and non-breeding seasons, respectively; P<0.01) and development (67.3±7.6% and 40.0±7.2% in the breeding and non-breeding seasons, respectively; P<0.01) rates of vitrified blastocysts decreased after 24h post-warming culture in the non-breeding season. These findings suggest that the reduced developmental competence of buffalo oocytes during the non-breeding season may also lead to lower blastocyst quality. This is in contrast to the evidence in cattle that embryo quality is mainly determined by culture conditions, whereas blastocyst production depends on oocyte quality.

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Effect of antioxidant ascorbic acid on in vitro maturation of Caprine oocytes under normal and elevated temperatures

Indian Journal of Animal Research ◽

10.18805/ijar.b-3616 ◽

2018 ◽

Author(s):

S.B. Khanday ◽

J.A. Ahmed ◽

N. Nashiruddullah ◽

U. Sharma and D. Chakraborty

Keyword(s):

Ascorbic Acid ◽

Heat Stress ◽

In Vitro Maturation ◽

Elevated Temperatures ◽

Developmental Competence ◽

Cumulus Expansion ◽

Nuclear Maturation ◽

Maturation Rate ◽

Metaphase Stage

The aim of the present study was to assess the effect of ascorbic acid on in vitro maturation of caprine oocytes under normal and elevated temperatures. Goat ovaries were collected at slaughter and both A and B grade cumulus-oocyte-complexes (COCs) were aspirated out and were matured in vitro under normal (38.5°C) and elevated temperatures (41°C). On the basis of cumulus expansion and nuclear maturation, the maturation competencewere compared with and without ascorbic acid supplementation (100 µM). Heat stress significantly (P£ 0.01) reduced cumulus expansion, maturation rate and lowered metaphase stage II of nuclear maturation. Ascorbic acid improved developmental competence of oocytes during heat stress (41 °C) and ascorbic acid supplemented COCs demonstrated significantly (P£ 0.05) higher maturation rates when compared to non-supplemented groups.

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Effects of gonadotrophins, growth hormone and prolactin on developmental competence of domestic cat oocytes matured in vitro

Reproduction Fertility and Development ◽

10.1071/rd9951061 ◽

1995 ◽

Vol 7 (5) ◽

pp. 1061 ◽

Cited By ~ 23

Author(s):

RD Schramm ◽

BD Bavister

Keyword(s):

Growth Hormone ◽

Calf Serum ◽

Cell Number ◽

Blastocyst Stage ◽

Domestic Cat ◽

Developmental Competence ◽

Developmental Potential ◽

Nuclear Maturation ◽

Morula Stage

Specific aims were to (1) examine the developmental capacity of felid oocytes matured in vitro and (2) determine the effects of gonadotrophins, growth hormone and prolactin on nuclear and cytoplasmic maturation oocytes in vitro. Oocytes were obtained from excised ovaries of 21 cats, and were matured for 45-46 h in modified CMRL-1066 culture medium (1 mM glutamine, 1 mM pyruvate and 20% bovine calf serum), with one of the following: (1) gonadotrophins (1.0 micrograms mL-1 hFSH+10 micrograms mL-1 hLH), (2) gonadotrophins+10 micrograms mL-1 growth hormone, (3) gonadotrophins+10 micrograms mL-1 prolactin, or (4) no hormones. Oocytes were inseminated with ejaculated cat sperm capacitated in TALP medium. Embryos were cultured in modified CMRL-1066 medium until developmental arrest, then stained with Hoechst 33342 to assess nuclear status or cell number. Gonadotrophins enhanced (P < or = 0.05) the incidence of nuclear maturation, but neither gonadotrophins, growth hormone nor prolactin improved fertilization or developmental potential of oocytes matured in vitro. Mean percentages of mature oocytes that were fertilized and cleaved to or beyond the 2, 4, 8 and 16-cell stages were 80, 77, 66, 42 and 24%, respectively. Three embryos progressed to 40-60 cells, but none developed a blastocoel. Thus, although gonadotrophins enhance nuclear maturation of oocytes in vitro, and mature oocytes are capable of fertilization and development to the morula stage, culture with growth hormone, prolactin or gonadotrophins during maturation in vitro does not enhance developmental competence or overcome the morula-to-blastocyst-stage block in development of domestic-cat oocytes matured in vitro.

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269 EFFECTS OF REPRODUCTIVE STATUS AND OOCYTE QUALITY ON THE DEVELOPMENTAL COMPETENCE OF OOCYTES FOLLOWING IN VITRO PRODUCTION IN DOMESTIC CAT

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab269 ◽

2006 ◽

Vol 18 (2) ◽

pp. 242

Author(s):

B. Agung ◽

P. Wongsrikeao ◽

H. Fuchieda ◽

T. Otoi

Keyword(s):

Reproductive Cycle ◽

Bovine Serum ◽

Cell Mass ◽

Cumulus Cell ◽

Oocyte Quality ◽

Domestic Cat ◽

Developmental Competence ◽

Human Menopausal Gonadotropin ◽

Significant Difference

A recent study showed that the developmental competence of cat oocytes after IVM and IVF was affected by the estrous cycle stage of the donor (Freistedt et al. 2001 Biol. Reprod. 65, 9-13). This study was conducted to examine the effect of the cat reproductive cycle stage and the oocyte quality on the developmental competence of the oocyte following IVF production. Cat ovaries were collected at veterinary clinics and stored at 4�C for 24 h. Based on the presence or absence of follicles and corpora lutea, the ovaries were classified into the luteal, follicular, or inactive stage. Cumulus oocyte complexes (COCs) from the different stage ovaries were separated at recovery into three ranks (A, B, and C) according to pigmentation, uniformity and smoothness of ooplasm, and amount of surrounding cumulus cell mass and were cultured separately in 100 �L drops of maturation medium (TCM-199), supplemented with 0.4% bovine serum albumin, 0.1 IU/mL (human menopausal gonadotropin (HMG), 10 IU/mL HCG, 1 �g/mL 17�-estradiol, and 100 �g/mL gentamicin) for 24-h at 38�C, 5% CO2 in air. After 24-h in vitro culture, the oocytes were transferred into 100 �L sperm microdrops of Brackett-Oliphant medium (2 � 106 sperm/mL) for fertilization and were co-incubated for 12 h. Subsequently, presumptive zygotes were transferred into a modified Earle's balanced salt solution (MK-1) supplemented with 4 mg/mL BSA and 50 �g/mL gentamicin. Three days after insemination, all embryos were transferred into culture medium of MK-1 supplemented with 5% (v/v) fetal bovine serum and 50 �g/mL gentamicin. The cleaved embryos were further cultured for 5 days to evaluate their ability to develop to the blastocyst stage. Data were analyzed by ANOVA. The percentages of COCs of A (35.5%, 31.8%, 27.7%), B (41.5%, 39.9%, 42.9%), and C (23.0%, 28.3%, 29.4%) ranks did not show significant differences (P > 0.05) among the reproductive stages of ovaries (luteal, follicular, and inactive, respectively). There were significant differences in the percentages of cleavage (P < 0.05) among the A, B, and C ranks of oocytes from ovaries classified as luteal and follicular stages (50%, 35%, 1.8%; 52%, 35%, 3.9%, respectively, P < 0.05). However, there were no significant differences between the A and B ranks of oocytes obtained from inactive stages of ovaries (47% vs. 44%, respectively; P > 0.05) but there was a significant difference for C rank (1.9%). The percentages of blastocyst formation were significantly different (P < 0.05) among the ranks of oocytes obtained from luteal, follicular, and inactive stages (27%, 14.6%, and 0% for A; 24%, 11.7%, and 0.6% for B; 35%, 21.6%, and 2.9% for C). However, there were no significant differences (P > 0.05) among the reproductive cycle stages of ovaries in the three oocyte ranks with respect to the percentages of cleavage and blastocyst formation.These results indicate that the reproductive stage of donor cat ovaries stored at 4�C for 24 h has no apparent effect on the developmental competence of the oocyte following IVF, but development is affected by oocyte quality.

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Effect of oxygen tension and antioxidants on the developmental competence of buffalo oocytes cultured in vitro

Veterinary World ◽

10.14202/vetworld.2021.78-84 ◽

2021 ◽

Vol 14 (1) ◽

pp. 78-84

Author(s):

Amro M. El-Sanea ◽

Ahmed Sabry S. Abdoon ◽

Omaima M. Kandil ◽

Nahed E. El-Toukhy ◽

Amal M. Abo El-maaty ◽

...

Keyword(s):

Ascorbic Acid ◽

Oxygen Tension ◽

Cumulus Cell ◽

Acid Group ◽

Developmental Competence ◽

Control Group ◽

In Vitro Production ◽

Nuclear Maturation ◽

Effect Of Oxygen

Aim: Oxidative stress (OS) is one of the major disruptors of oocyte developmental competence, which appears due to the imbalance between the production and neutralization of reactive oxygen species (ROS). Materials and Methods: In Experiment 1, buffalo oocytes were in vitro matured, fertilized, and cultured at 38.5°C under 5% CO2 + 20% O2 in standard CO2 incubator (OS) or under 5% O2 + 5% CO2 + 90% N2 (Multi-gas incubator, low O2). In Experiment 2, buffalo cumulus oocytes complexes (COCs) were matured in Basic maturation medium (BMM) composed of TCM199+ 10% FCS+ 10 μg/ml FSH+ 50 μg/ml gentamicin (control group) or in BMM supplemented with 50 μM ascorbic acid (ascorbic acid group) or 3.0 mM glutathione (glutathione group) or 10-5 M melatonin (melatonin group) and cultured at 38.5°C under 20% O2 for 24 h. Matured buffalo oocytes in control, ascorbic acid, or melatonin groups were fertilized and zygotes were cultured for 8 days under the same conditions. Results: In both experiments, maturation, cleavage, and blastocyst rates were recorded. Results showed that culture of buffalo oocytes under low O2 (5% O2) significantly increased maturation, cleavage, and blastocyst rates (p<0.05). Meanwhile, under 20% O2, addition of 10-5 M melatonin or 50 μM ascorbic acid to in vitro maturation (IVM) medium significantly improved cumulus cell expansion, nuclear maturation rates of buffalo oocytes (p<0.05), and increased cleavage and blastocyst rates (p<0.05). Conclusion: About 5% O2 is the optimum condition for in vitro production of buffalo embryos, and addition of 10-5 M melatonin to IVM medium for oocytes cultured under 20% O2 could alleviate the adverse effect of high oxygen tension and increased embryo yield.

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