scholarly journals Graft site and gonadotrophin stimulation influences the number and quality of oocytes from murine ovarian tissue grafts

Reproduction ◽  
2006 ◽  
Vol 131 (5) ◽  
pp. 851-859 ◽  
Author(s):  
Hsiao Yun Yang ◽  
Shae-Lee Cox ◽  
Graham Jenkin ◽  
Jock Findlay ◽  
Alan Trounson ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Germinal Vesicle ◽  
Fertilization Rate ◽  
Ovarian Tissue Cryopreservation ◽  
Control Group ◽  
Developmental Potential ◽  
Oocyte Yield ◽  
Tissue Grafts

Ovarian tissue cryopreservation and subsequent transplantation can restore fertility in cancer patients. This study used a mouse ovarian grafting model to investigate whether the graft site (bursal cavity, the kidney capsule or subcutaneous) influences the number, fertilization rate and developmental potential of oocytes recovered from grafts and whether using a standard gonadotrophin stimulation protocol would increase oocyte yield from the grafts. Mouse ovarian tissue was grafted into four week old mice and collected three weeks later. Graft recipients were treated either with or without exogenous gonadotrophin stimulation prior to graft collection. Grafted ovaries yielded oocytes that were either at the germinal vesicle (GV) stage or mature metaphase II (MII) stage at collection. These GV oocytes were matured beforein vitrofertilization (IVF), while the MII oocytes underwent IVF immediately. Oocytes collected from the oviducts of non-grafted superovulated mice of the same age served as controls. Two-cell embryos were transferred to pseudopregnant recipients and recovered at day 15 of gestation or left to go to term. Graft retrieval and the number of oocytes from each graft were lowest from the subcutaneous graft site. The number of two-cell embryos produced was significantly higher for oocytes from the grafts to the bursa as compared with the other sites. All graft sites gave rise to embryos with comparable implantation rates and developmental potential to fetuses and offspring following transfer. However, the oocytes from grafted ovaries had a significantly lower developmental potential when compared with the control group. Stimulation with exogenous gonadotrophins did not significantly increase oocyte yield from grafted ovaries but did enhance oocyte maturation and development. In conclusion, graft site affects the number and quality of oocytes produced from ovarian grafts.

scholarly journals Live offspring from vitrified blastocysts derived from fresh and cryopreserved ovarian tissue grafts of adult mice

Reproduction ◽  
2009 ◽  
Vol 138 (3) ◽  
pp. 527-535 ◽  
Author(s):  
Xiaoqian Wang ◽  
Sally Catt ◽  
Mulyoto Pangestu ◽  
Peter Temple-Smith
Keyword(s):  
Cancer Patients ◽  
Oocyte Maturation ◽  
Ovarian Tissue ◽  
Subsequent Development ◽  
Cumulus Cells ◽  
Ovarian Tissue Cryopreservation ◽  
Blastocyst Formation ◽  
Adult Mice ◽  
Tissue Grafts

Ovarian tissue cryopreservation and transplantation can be used to preserve fertility for cancer patients. In this study, we assessed the viability and function of ovarian tissue from adult mice that was cryopreserved by solid surface vitrification or traditional slow-cooling using variousin vitroandin vivotechniques, including allotransplantation,in vitrooocyte maturation, embryo culturein vitro, blastocyst cryopreservation, embryo transfer, and development. The importance of cumulus cells for oocyte maturation, fertilization, and embryo development was investigated. Graft recovery, follicle survival, and oocyte retrieval was similar in control, vitrified, and slow-cooled groups. High rates of oocyte maturation, cleavage, and blastocyst formation were achieved, with no significant differences between the control, vitrified or slow-cooled ovarian tissue grafts. The presence of cumulus cells was important for oocyte maturation, fertilization, and subsequent development. Cumulus–oocyte complexes with no surrounding cumulus cells (N-COCs) or with an incomplete layer (P-COCs) had significantly lower rates of oocyte maturation and blastocyst formation than cumulus–oocyte complexes with at least one complete layer of cumulus cells (F-COCs; maturation rate: 63, 78 vs 94%; blastocyst rate: 29, 49 vs 80%). Live births were achieved using vitrified blastocysts derived from oocytes taken from vitrified and slow-cooled ovarian tissue heterotypic allografts. Successful production of healthy offspring from these vitrified blastocysts suggests that this technique should be considered as a useful stage to pause in the assisted reproduction pathway. This provides an alternative protocol for restoring fertility and offering cancer patients a better indication of their chances of pregnancy and live birth.

Cycloheximide speeds the porcine oocyte nuclear kinetics and retains embryonic developmental potential in the presence of gonadotrophins

2010 ◽  
Vol 90 (2) ◽  
pp. 189-196
Author(s):  
X -L. Sun ◽  
W -Z. Ma ◽  
Y -B. Zhu ◽  
Z -H. Wu ◽  
L. An ◽  
...  
Keyword(s):  
Embryo Development ◽  
Germinal Vesicle ◽  
Chorionic Gonadotrophin ◽  
Developmental Competence ◽  
Electrical Activation ◽  
Control Group ◽  
Germinal Vesicle Breakdown ◽  
Developmental Potential ◽  
Oocyte Meiosis

Animal embryo engineering requires large amounts of synchronized mature oocytes in vitro. However, porcine cumulus-oocyte complexes aspirated from 3-8 mm follicles are at different germinal vesicle stages. They reach metaphase II stages asynchronously when cultured in vitro. In this study, we examined the effects of pretreatment with or without cycloheximide (CHX), equine chorionic gonadotrophin (eCG), human chorionic gonadotrophin (hCG), and their combinations on meiotic synchronization and the developmental competence of porcine oocytes in vitro following electrical activation. The COCs were pretreated for 12 h with either control medium (TCM 199), CHX (TCM 199 + CHX), eCG/hCG (TCM 199 + eCG/hCG) or eCG/hCG + CHX (TCM 199 + CHX + eCG/hCG), and then cultured for up to 32 h with TCM199 + eCG/hCG. After 12 h pretreatment, the rates of germinal vesicle breakdown (GVBD) were lower (P < 0.05) in the CHX (8.4%) and eCG/hCG + CHX (1.5%) groups compared with control (55.4%) and eCG/hCG (27.2%) groups. After removal of CHX and culture for an additional 12 h in vitro, the majority of the oocytes were synchronized at the GVBD stage in CHX (75.6%) and eCG/hCG + CHX (65.0%) groups. At additional 32 h of culture, the rate of oocytes in metaphase II in eCG/hCG + CHX group (68.3%) was significantly (P < 0.05) higher than the eCG/hCG group (54.8%), but did not differ from other groups (control: 61.3%, CHX: 58.8%). After electrical activation, the cleavage and blastocyst formation rates in the CHX group (80.3%; 19.5%) were significantly (P < 0.05) lower than those in the control group (95.5%; 45.3%), while no difference was found between eCG/hCG + CHX (82.2%; 34.4%) and control groups. Our data, hence, demonstrate pretreatment with CHX hastened nuclear kinetics of porcine oocytes cultured in vitro; however, embryo development potential was retained only when gonadotrophins is present in the in vitro maturation (IVM) medium. Thus, CHX should be used in the two-step culture systems in combination with gonadotrophins. Key words: Oocyte meiosis, synchronization, cycloheximide, embryo development, pig

318 EFFECT OF SPERM PRETREATMENT WITH ISOBUTYLMETHYLXANTHINE AND METHYL-β-CYCLODEXTRIN ON THE EFFICIENCY OF BOVINE INTRACYTOPLASMIC SPERM INJECTION

2015 ◽  
Vol 27 (1) ◽  
pp. 248
Author(s):  
L. M. Aguila ◽  
M. E. Arias ◽  
R. S. Sanchez ◽  
T. C. Vargas ◽  
F. A. Zambrano ◽  
...  
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Defined Medium ◽  
Fertilization Rate ◽  
Vehicle Group ◽  
Control Group ◽  
Sperm Capacitation ◽  
Chi Square ◽  
Developmental Potential ◽  
Pronuclear Formation

The efficiency of intracytoplasmic sperm injection (ICSI) in bovines is lower than in other species. We propose that in vitro sperm capacitation could optimize the ICSI in cattle. The aim was to evaluate the effects of isobutylmethylxanthine (IBMX) and methyl-β-cyclodextrin (MβCD) on the sperm capacitation and in vitro development of embryos generated by ICSI. Frozen-thawed spermatozoa (3–5 × 106 cells mL–1) were pre-incubated for 2 h at 38.5°C, 5% CO2 in defined medium (Sp-TLP/PVA) supplemented with MβCD (1 mM) or IBMX (0.4 mM) (capacitating conditions). The untreated control group (UTG; not supplemented) and vehicle group (VG) were incubated for 2 h. The non-capacitating control group (NCG) was not supplemented (neither vehicle nor IBMX or MβCD) and not incubated. The sperm viability and capacitation {intracellular calcium [Ca2+]i, plasma membrane fluidity (PMF), and acrosomal reaction} were evaluated by flow cytometry (n = 3 biological replicates). For the ICSI procedure, only motile spermatozoa were selected. After ICSI, oocytes were activated with ionomycin + cycloheximide. Culture was performed at 38.5°C, 5% CO2, 5% O2, 90% N2, saturation humidity in KSOM base medium. Data were analysed by ANOVA and Scheffe's test. Pronuclear formation was evaluated by a chi-square test with Bonferroni's correction. Significance was set at P < 0.05. Pretreated spermatozoa showed lower (P < 0.05) viability (49 and 67% for IBMX and MβCD, respectively) compared with the NCG (89%), UTG (80%), and VG (78%). The [Ca2+]I analysed by median fluorescence intensity (MFI) was lower (P < 0.05) in NCG (117 MFI) with respect to UTG (127 MFI), VG (124 MFI), IBMX (126 MFI), and MβCD (131 MFI). The PMF increased (P < 0.05) with IBMX (115 MFI) and MβCD (106 MFI) compared with NCG (70 MFI), UTG (89 MFI), and VG (65 MFI). Acrosome reaction improved with capacitating treatments with respect to both control groups (16, 23, 8, 4, and 3% for IBMX, MβCD, UTG, VG, and NCG, respectively). Analysis of capacitating v. non-capacitating conditions on ICSI efficiency revealed that the fertilization rate, assessed by pronuclear formation, was higher (P < 0.05) in ICSI-MβCD (76%; n = 46) compared with ICSI-IBMX (55%; n = 53) and ICSI-NCG (50%; n = 44). Nevertheless, there were no differences among groups in cleavage (Day 3): 85, 86, and 84% and blastocyst rates (Day 8): 19, 25, and 18% for ICSI-IBMX (n = 8), ICSI-MβCD (n = 7), and ICSI-NCG (n = 7), respectively. The parthenogenetic and sham injection groups yielded a lower rate of cleavage (73 and 53%, respectively) and blastocyst (13% and 10%, respectively). The results demonstrated an improvement of the fertilization rate of bovine embryos generated by ICSI using sperm capacitated by MβCD pretreatment. However, more studies are necessary to improve in vitro developmental potential of these embryos to the blastocyst stage.Frigorífico Temuco and funding support from FONDECYT 1120241 CONICYT-Chile are gratefully acknowledged.

Oocytes recovered after ovarian tissue slow freezing have impaired H2AX phosphorylation and functional competence

2015 ◽  
Vol 27 (8) ◽  
pp. 1242
Author(s):  
Sam Sudhakaran ◽  
Shubhashree Uppangala ◽  
Sujith Raj Salian ◽  
Sachin D. Honguntikar ◽  
Ramya Nair ◽  
...  
Keyword(s):  
Ovarian Tissue ◽  
Germinal Vesicle ◽  
Slow Freezing ◽  
Ovarian Tissue Cryopreservation ◽  
Strontium Chloride ◽  
H2ax Phosphorylation ◽  
Germinal Vesicle Breakdown ◽  
Functional Competence ◽  
Dna Strand

It has been shown that oocytes isolated from ovarian tissue cryopreservation acquire DNA damage during the process of freeze–thawing. Using a mouse model, here we have investigated the functional competence and phosphorylation of H2AX (γ-H2AX) in germinal vesicle (GV) and parthenogenetically activated oocytes derived from conventional ovarian tissue slow freezing and vitrification techniques. The number of GV-stage oocytes with γ-H2AX foci was not significantly different between the slow-freezing and vitrification groups. Although the in vitro maturation (IVM) potential of GV oocytes in the slow-freezing group showed a significant delay (P < 0.0001) in the process of germinal vesicle breakdown, no difference in the maturation rate was observed between the two protocols. Nevertheless, parthenogenetic activation of IVM oocytes using strontium chloride showed a significantly lower activation rate in the slow-freezing group compared with the vitrification (P < 0.05) and control (P < 0.01) groups. Importantly, H2AX phosphorylation was significantly perturbed in the slow-freezing group in comparison to the control (P < 0.05). Therefore, we conclude that impaired sensing of DNA strand breaks and repair processes are associated with the reduced functional competence of the oocytes recovered from the slow-freezing group, which may have a significant impact on the reproductive outcome.

Developmental potential of in vitro or in vivo matured oocytes

Zygote ◽  
2013 ◽  
Vol 23 (1) ◽  
pp. 93-98 ◽  
Author(s):  
Diego D. Alcoba ◽  
Anita M. Pimentel ◽  
Ilma S. Brum ◽  
Helena E. Corleta
Keyword(s):  
Germinal Vesicle ◽  
Fertilization Rate ◽  
Cleavage Rate ◽  
Suitable Alternative ◽  
Developmental Potential ◽  
Immature Oocytes ◽  
Prophase I ◽  
Intra Cytoplasmic Sperm Injection

SummaryThis study compared the embryological features of mature and immature oocytes (different stages) collected from stimulated cycles of in vitro fertilization (IVF). Immature oocytes were identified, classified as PI (prophase I – germinal vesicle, GV) or MI (metaphase I), were matured in vitro and fertilized using the intra-cytoplasmic sperm injection (ICSI) technique. Fertilization potential, cleavage, and subsequent transfer/cryopreservation of the embryos derived from these in vitro matured oocytes were compared with those of in vivo matured oocytes (collected at the MII stage). The characteristics of embryos derived from gametes recovered in the MI and MII stages were similar. The fertilization rate of immature oocytes recovered in PI was significantly lower than that of MII oocytes (P = 0.031), and the cleavage rate of the PI group was also lower than that of the MI (P = 0.004) and MII (P < 0.001) groups. In vitro maturation of MI oocytes is a suitable alternative when immature oocytes are recovered, as their characteristics and development are similar to those of in vivo matured oocytes. Optimization of outcomes for PI oocytes will require development of techniques that can distinguish which of these gametes will mature and fertilize.

scholarly journals 221.Effects of exogenous gonadotrophin stimulation on ovarian tissue grafts in the mouse

2004 ◽  
Vol 16 (9) ◽  
pp. 221
Author(s):  
H. Yang ◽  
S. Cox ◽  
J. Shaw ◽  
G. Jenkin
Keyword(s):  
Subcutaneous Tissue ◽  
Ovarian Tissue ◽  
Control Group ◽  
Control Groups ◽  
Hybrid Strain ◽  
Tissue Grafts ◽  
And Control

Ovarian tissue grafts commonly contain only limited numbers of follicles. The functional life span and ability to retrieve as many mature oocytes as possible from ovarian grafts is important when grafting is used to restore fertility. This study aimed to determine whether ovarian grafts responded to exogenous hormones in a similar manner to that of in situ ovaries. Ovaries of C57BlxCBA F1 mice were cut in half and grafted to one of three different graft sites in females of the same F1 line; bursal capsule (BC, n = 12), kidney capsule (KC, n = 6), subcutaneous tissue (SC, n = 24). Three weeks after grafting, half of the graft recipients in each group were treated with 5IU PMSG followed by 5IU hCG 48 hours later. Oocytes were collected directly from the grafted ovaries 10 hours after the hCG injection and fertilized in vitro. Oocytes from the ovaries of superovulated normal mice (n = 4) of the same hybrid strain were used as controls. Two-cell embryos were transferred to pseudopregnant recipients and collected at day 15 of gestation or the animals were allowed to go to term. Mature fertilisable MII oocytes were retrieved from stimulated grafts from all graft sites, however, the number (BC 9, KC 5, SC 2 oocytes per ovary) and proportion of two-cell embryos in each grafted group (BC 52%, KC 32%, SC 32%) was significantly (P < 0.05) lower than in the in vivo matured control (16 oocytes, 85% two-cell). The fetal and placental weights of fetuses produced from graft-derived oocytes were not significantly different to the control group. Phenotypically normal pups were born in each of the graft and control groups. In conclusion, ovarian grafts treated with exogenous gonadotrophins produce significantly fewer mature oocytes and two cell embryos compared to in situ ovaries. Work supported by ARC and NIH RFA.

126 EFFECTIVE METHOD FOR IN VITRO CULTURE OF CRYOPRESERVED OVINE OVARIAN TISSUE

2016 ◽  
Vol 28 (2) ◽  
pp. 193
Author(s):  
A. Seisenbayeva ◽  
Y. Toishibekov ◽  
U. Iglmanov ◽  
B. Valiyeva ◽  
B. Katubayeva
Keyword(s):  
In Vitro Culture ◽  
Room Temperature ◽  
Culture Media ◽  
Ovarian Tissue ◽  
Farm Animals ◽  
Penicillin G ◽  
Ovarian Tissue Cryopreservation ◽  
Control Group ◽  
Primordial Follicles

Today, ovarian tissue cryopreservation is used for preserving the reproductive function of women, as well as the genetic material of rare and endangered species or domestic animals breeds. In the last 20 years genetic diversity of farm animals breeds suffered considerable losses in Kazakhstan; therefore, genetic preservation of valuable local breeds is desirable. The aim of this study was to compare the effectiveness of different in vitro culture media on morphology of ovine ovarian tissue cryopreserved by a slow-freezing protocol with 1.5 M dimethyl sulfoxide (DMSO). Ovaries were collected from indigenous Chuyi breed and immediately transported to the laboratory at 30°C within 1 h. Ovaries were rinsed several times in PBS supplemented with antibiotics (75 mg L–1 of penicillin-G, 50 mg L–1 of streptomycin sulfate). In Hepes-buffered medium 199, halved and the medulla removed with curved iris. Using a scalpel, the cortex was cut into 5- × 3- × 1-mm strips. Ovarian strips were equilibrated sequentially in freezing medium containing 0.25, 0.75, and 1.5 M DMSO with 0.5 M sucrose (5 min each). Then, ovarian strips were frozen in plastic straws using a programmable freezer Planer Kryo-360 3,3 (Planer, UK) and cooled as follows: stabilised at 20°C for 5 min, cooled from 20°C to –70°C at 5°C min–1, seeded to the temperature –7°C, cooled again to –30°C at 0.3°C min–1, cooled to –150°C at 35°C min–1, and finally plunged into liquid nitrogen and stored for 10 days. The straws were thawed at room temperature for 1 min, and then immersed in a water bath at 37°C for 2 min, warmed at room temperature with Dulbecco’s PBS (DPBS), supplemented with 10% FCS and 0.75 M sucrose (15 min), then DPBS + 10% FCS (30 min), and finally placed in the culture media for 10 min. Fresh and frozen tissue pieces were randomly distributed into 12 groups for further culture: 1) TCM 199 + 10% FBS; 2) TCM 199 + 10% native ovine serum (NOS); 3) TCM-Hepes + 10% FBS; 4) TCM-Hepes + 10% NOS; 5) DMEM + 10% FBS; 6) DMEM + 10% NOS; with and without 7.5 mg mL–1 of FSH. After 7 days of culture, the effects of different culture media on ovarian tissue morphology was evaluated by light microscopy after hematoxylin and eosin staining of tissue sections. The best result was observed when frozen ovarian tissue was cultured in the presence of FSH. The best result was observed in group 3 and 4 with FSH. The percentages of normal primordial, primary, and preantral follicles were: 1) TCM 199 + 10% FBS + FSH = 53.5 ± 3.1, 39.7 ± 3.8a, 28.5 ± 3.2; 2) TCM 199 + 10% NOS + FSH = 49.4 ± 2.3a, 36.7 ± 3.3a, 25.3 ± 4.1; 3) TCM-Hepes + 10% FBS + FSH = 66.3 ± 2.5, 45.7 ± 3.9, 35.1 ± 3.8; 4) TCM-Hepes + 10% NOS + FSH = 86.5 ± 3.8b, 75.4 ± 4.2b, 45.7 ± 3.5; 5) DMEM + 10% FBS + FSH = 42.1 ± 3.5a, 33.7 ± 2.9a, 21.3 ± 4.9, 20.7 ± 3.9; 6) DMEM + 10% NOS + FSH = 41.3 ± 3.9a, 32.9 ± 2.5a; control group = 98.2 ± 1.1, 93.7 ± 1.7, 90.3 ± 1.9 (ab, P < 0.01). The majority of follicles in groups without FSH were degenerated. In group 4) TCM-Hepes + 10% NOS without FSH, a damaged structure of primordial follicles was observed.

41 DEVELOPMENTAL POTENTIAL OF BUFFALO OOCYTES VITRIFIED AT THE GERMINAL VESICLE STAGE: EFFECTS OF DIFFERENT CRYOPROTECTANT COMBINATIONS AND CRYODEVICES

2016 ◽  
Vol 28 (2) ◽  
pp. 150
Author(s):  
A. S. El-Shalofy ◽  
A. R. Moawad ◽  
G. M. Darwish ◽  
S. T. Ismail ◽  
A. B. Badawy
Keyword(s):  
Solid Surface ◽  
Germinal Vesicle ◽  
Subsequent Development ◽  
The Other ◽  
Control Group ◽  
Developmental Potential ◽  
High Frequencies ◽  
Nuclear Maturation ◽  
Maturation Rate

The cryopreservation of immature oocytes would generate a readily available, nonseasonal source of female gametes for both research and reproduction. In domestic animals, the most promising results in the field of oocyte cryopreservation have been reported in cattle, and a few experiments have been conducted on buffalo. The aim of the present study was to compare the effects of different cryoprotectant combinations and different cryodevices on viability and subsequent development of buffalo oocytes vitrified at the germinal vesicle stage. Cumulus-oocyte complexes obtained at slaughter from mature buffalos were vitrified by using either straw or open pulled-straw or solid surface vitrification (SSV) in a solution composed of either 20% ethylene glycol (EG) + 20% glycerol or 20% EG + 20% dimethylsulfoxide (DMSO). Following vitrification and warming, viable oocytes were matured in vitro for 22 h. Matured oocytes were either evaluated for nuclear maturation or fertilized and cultured in vitro for 7 days. Recovery rate was significantly higher (P < 0.05) in the oocytes vitrified by straw in 20% EG + 20% glycerol (92.6%) as compared with the other groups. Percentages of viable oocytes were significantly higher (P < 0.05) in the oocytes vitrified in 20% EG + 20% DMSO using SSV (95.7%) than those in the other groups (from 80 to 88.0%). Among the vitrified groups, the highest maturation rate was achieved in SSV with 20% EG + 20% DMSO group (56.7%). This value was comparable with those in the control group (62.1%). After IVF and embryo culture, the highest cleavage and blastocyst rates were obtained in SSV with 20% EG + 20% DMSO group (35.7 and 21.4%, respectively), and these values were nearly similar to those in the control group (38.7 and 25.8%, respectively). Vitrification of oocytes by straw or open pulled-straw resulted in significantly lower (P < 0.05) blastocyst rates (2.6 and 11.5%) as compared with the control. In conclusion, buffalo oocytes vitrified at the germinal vesicle stage can be matured, fertilized, and developed in vitro and produce high frequencies of blastocyst embryos. Solid surface vitrification may be superior to straw and open pulled-straw in vitrification of immature buffalo oocytes because this technique results in higher survival and embryo development rates.

scholarly journals Effects of prematuration culture with a phosphodiesterase-3 inhibitor on oocyte morphology and embryo quality in in vitro maturation

2021 ◽  
Vol 48 (4) ◽  
pp. 352-361
Author(s):  
Mohammed Ashraf Cheruveetil ◽  
Prasanna Kumar Shetty ◽  
Kamini A Rao ◽  
Arya Rajendran ◽  
Muhammed Asif
Keyword(s):  
In Vitro Maturation ◽  
Embryo Quality ◽  
Germinal Vesicle ◽  
Control Group ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Phosphodiesterase 3 ◽  
The Impact ◽  
Experimental Group

Objective: The study assessed the developmental potential of germinal vesicle (GV) oocytes subjected to in vitro maturation (IVM) after prematuration culture with cilostamide (a phosphodiesterase-3 inhibitor) and the impact of cilostamide exposure on the morphology of meiosis II (MII) oocytes and subsequent embryo quality. Methods: In total, 994 oocytes were collected from 63 patients. Among 307 GV oocytes, 140 oocytes were selected for the experimental group and 130 oocytes for the control group. The denuded GV-stage oocytes were cultured for 6 hours with cilostamide in the experimental group and without cilostamide in the control group. After 6 hours, the oocytes in the experimental group were washed and transferred to fresh IVM medium. The maturational status of the oocytes in both groups was examined at 26, 36, and 48 hours. Fertilization was assessed at 18 hours post-intracytoplasmic sperm injection. Embryo quality was assessed on days 3 and 5.Results: In total, 92.1% of the oocytes remained in the GV stage, while 6.4% converted to the MI stage (p<0.01) after cilostamide exposure. In both groups, more MII oocytes were observed at 36 hours (25.8% vs. 21.5%) than at 26 hours (10.8% vs. 14.6%) and 48 hours (13% vs. 7.9%) (p>0.05). With the advenet of cilostamide, blastocyst quality was better in the experimental group than in the control group (p<0.05). Conclusions: Cilostamide effectively blocked nuclear maturation and promoted cytoplasmic growth. Prematuration culture with cilostamide enabled synchronization between cytoplasmic and nuclear maturity, resulting in better blastocyst outcomes.

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