P–045 A retrospective study on Hyaluronic acid binding sperm selection in ICSI cycles

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
J Y Lo ◽  
S T Tee
Keyword(s):  
Pregnancy Rate ◽  
Fertilization Rate ◽  
Mature Sperm ◽  
Utilization Rate ◽  
Dna Integrity ◽  
Sperm Selection ◽  
Miscarriage Rate ◽  
Icsi Cycle ◽  
Significant Difference

Abstract Study question Does Hyaluronic acid(HA) binding sperm selection prior ICSI produce better outcome compare to PVP? Summary answer No significant difference in fertilization rate, blastocyst utilization rate, pregnancy rate and miscarriage rate in HA vs PVP group. What is known already HA is natural, non-sulphated glycosaminoglycan secretion that abundantly found in the cervical mucus and the cumulus oophorus complex (COC). In in-vivo, HA binding sites on the sperm plasma membrane indicate sperm maturity, mature sperm reaching HA-coated COC can bind and digest HA and subsequent hyperactivation further facilitate fertilization. In IVF, the use of HA tries to mimic in-vivo to select mature sperm with high DNA integrity prior to ICSI. In HA, movement of mature sperm is ‘slowed’ thus allow the selection of sperm to be used in ICSI. Sperm immaturity is known to associated with aneuploidy incidence in the embryos. Study design, size, duration ICSI cycle using HA (N = 83) was adapted from January to December 2020 while ICSI cycle using PVP (N = 133) was adapted from January 2018 to December 2019. Mean age of patient were 35.64±4.33 vs 34.15±4.75 for HA vs PVP group respectively. Fertilization rate, blastocyst utilization rate, pregnancy rate and miscarriage rate were recorded. This study included all ICSI cycle and both frozen and fresh embryo transfer data. Surgical retrieved sperm was excluded from this study. Participants/materials, setting, methods A 1.5ul of treated spermatozoa suspension was connected with a pipette tip to a 5ul droplet of fresh holding medium (Cook Gamete Buffer). Simultaneously, a 5ul droplet of HA medium (Origio SpermSlow) was connected to the 5ulL droplet of holding medium in a ICSI dish (Sparmed Oosafe) covered with oil (Vitrolife Ovoil). Sperm which were ‘slowed’ in the Sperm slow droplets with normal morphology according to WHO 2010 guideline were selected for ICSI at 400X. Main results and the role of chance The fertilization rate of the HA vs PVP- binding sperm are 68.6% vs 66.2%. As for the blastocyst utilisation rate is 61.6% vs 73.22% for HA vs PVP- binding sperm group. Pregnancy and miscarriage rate for HA vs PVP are 42.3% vs 51.5% and 19.4% vs 26.2% respectively. There was no significant difference in the fertilization rates, blastocysts utilisation rate, pregnancy rate and miscarriage rate between the HA vs PVP- binding sperm groups (P < 0.05). However, a trend of higher pregnancy (51.5% vs 42.3%; P = 0.279) and miscarriage rate were observed in the PVP group (26% vs 19%; P = 0.545) as compared to the HA group, but the difference was not statistically significant. The reason behind this might be the HA assist to select the mature sperm with higher DNA integrity and low frequency of chromosomal aneuploidies which contribute to the lower miscarriage rate in the study. Limitations, reasons for caution This is a retrospective study on HA binding sperm selection vs PVP prior to ICSI. Further research which includes a large number of RCT sample size should be warranted. Wider implications of the findings: The HA- sperm binding selection ICSI might only be beneficial to certain group of patients (high- DNA fragmentation sperm). A larger RCT study may be necessary to establish a relationship between HA-sperm binding selection vs aneuploidy rate via PGT analysis. Trial registration number Not applicable

P–131 Significance of the phenomenon of blastomere exclusion from compaction: Its relation to irregular cleavage, blastocyst development rate, and pregnancy rate

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
S Watanabe ◽  
M Tomida ◽  
S Suzuki ◽  
Y Matsuda ◽  
K Yoshikai ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Chromosome Analysis ◽  
Development Rate ◽  
Pregnancy Rates ◽  
Blastocyst Transfer ◽  
Blastocyst Development ◽  
Miscarriage Rate ◽  
Single Blastocyst Transfer ◽  
Significant Difference ◽  
Irregular Cleavage

Abstract Study question When does blastomere exclusion from compaction increase and what effect does it have on the embryo? Summary answer More blastomere were excluded from compaction in embryos with irregular cleavage, resulting in lower blastocyst development rates, but no decrease in pregnancy rates at transfer. What is known already It has been reported that many of the chromosome analysis results of blastomere excluded from compaction were aneuploid, and pointed out that this exclusion may be related to the repair of blastocyst euploidy, but the effect of the number of excluded blastomere has not been reported. Study design, size, duration This is a retrospective study of 578 embryos that developed into morula with time-lapse monitoring by EmbryoScope (Vitrolife) in 2018–2019. Participants/materials, setting, methods The target embryos were classified into two groups: embryos with normal first and second cleavage (normal cleavage group) and embryos with irregular cleavage (dynamics of one cell dividing into three or more cells), called “direct cleavage”, at either cleavage (DC group), and the number of blastomere excluded from compaction during morula formation was recorded and compared. The blastocyst development rate and single blastocyst transfer pregnancy rates of the two groups were compared. Main results and the role of chance There are 286 in the normal cleavage group and 292 in the DC group. The mean number of excluded blastomere was 0.76 and 3.55, respectively, which was significantly higher in the DC group (P < 0.01). Good blastocyst (Gardner classification 4 or higher) development rate was 84.5% (239/283) and 65.8% (181/275), respectively, and high grade blastocyst (Gardner classification BB or higher) development rate was 43.9% (105/239) and 14.9% (27/181) of them, both significantly higher in the normal cleavage group (P < 0.01). The single blastocyst transfer pregnancy rates were 31.6% (25/79) and 32.4% (11/34), and the miscarriage rates were 24.0% (6/25) and 27.3% (3/11), respectively, neither was there a significant difference between the two groups. So, direct cleavage increased the number of blastomere excluded from compaction, decreased the rate of morula to good blastocyst development and reduced blastocyst grade, but did not affect blastocyst transfer pregnancy rate and miscarriage rate. Limitations, reasons for caution Please note that all target embryos must have developed into morula or larger (embryos that did not develop into morula will not be included in the study). Wider implications of the findings: Severe chromosomal aberrant blastomeres formed by direct cleavage were excluded from compaction, and the blastocyst development rate decreased due to a decrease in the amount of viable cells, but it is suggested that this blastomere exclusion mechanism is not related to euploidy after blastocyst development. Trial registration number Not applicable

scholarly journals Bovine sperm selection procedure prior to cryopreservation for improvement of post-thawed semen quality and fertility

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
MariaPortia B. Nagata ◽  
Junki Egashira ◽  
Naoto Katafuchi ◽  
Kenji Endo ◽  
Kazuko Ogata ◽  
...  
Keyword(s):  
Artificial Insemination ◽  
Structural Integrity ◽  
Semen Quality ◽  
Selection Procedure ◽  
Dna Integrity ◽  
Sperm Selection ◽  
Animal Reproduction ◽  
Fertility Variation ◽  
The Individual

Abstract Background The application of cryopreservation and artificial insemination technology have contributed to the advancement of animal reproduction. However, a substantial proportion of spermatozoa undergoes alterations and loses their fertility during cryopreservation, rendering the frozen-thawed semen impractical for routine use. Cryopreservation is known to reduce sperm lifespan and fertility. Variation in cryosurvival of spermatozoa from different sires and even with the individual sire is common in artificial insemination (AI) centers. Our goal is to improve post-thawed semen quality by optimization of cryopreservation technique through sperm selection prior to cryopreservation process. Results Our strategy of sperm selection based on rheotaxis and thermotaxis (SSRT) on macrosale in a rotating fluid flow demonstrated the ability to maintain the original pre-freezing structural integrity, viability and biological function related to fertilization competence. This strategy has a positive effect on the cryosurvival and fertilizing abilities of spermatozoa as supported by the improvement on pregnancy rate of Japanese Black heifers and Holstein repeat breeders. This technique protected further sublethal damage to bovine spermatozoa (higher % cryosurvival than the control) and resulted in the improvement of DNA integrity. Prefreeze selected spermatozoa demonstrated slower and controlled capacitation than unprocessed control which is thought to be related to sperm longevity and consequently to appropriate timing during in vivo fertilization. Conclusions These results provide solid evidence that improvement of post-thawed semen quality by SSRT method is beneficial in terms of cryosurvival, longevity of post-thawed sperm, and optimization of in vivo fertilization, embryo development and calving as supported by the favorable results of field fertility study.

15 QUANTIFICATION OF BULL SPERM TRAITS AS ASSESSED BY COMPUTER-ASSISTED SEMEN ANALYSIS AND THE RELATIONSHIP TO PREGNANCY RATE FOLLOWING CONTROLLED BREEDING

2017 ◽  
Vol 29 (1) ◽  
pp. 115
Author(s):  
M. L. Mphaphathi ◽  
M. M. Seshoka ◽  
F. V. Ramukhithi ◽  
Z. C. Raphalalani ◽  
T. R. Netshirovha ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Pregnancy Rate ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Bull Sperm ◽  
Sperm Traits

The bull’s contribution through artificial insemination to reproductive efficiency is of great biological importance. The objectives were (1) to compare the oestrous synchronization response of Bonsmara and Nguni cows; and (2) to find the relationship between cow’s conception rate (in vivo and in vitro fertilization) and bull sperm motility rate assessed by computer-assisted semen analysis (CASA) following AI. For the in vivo sperm fertility test, 100 Bonsmara and 482 Nguni cows were randomly selected and subjected to oestrous synchronization protocol and AI with frozen–thawed assessed semen by CASA before AI. Briefly at Day 0, cows were inserted with an intravaginal CIDR® (1.9 g), which was removed on Day 7. Prostaglandin was then administered (2 mL) on Day 8 and a heatmount detector was placed on the hindquarter of each cow. For the in vitro sperm fertility test, collected oocytes from slaughterhouse were in vitro matured (n = 360) and in vitro fertilized (sperm/mL) in 100-µL droplets (final volume) of BO-IVF medium per treatment bulls (Bonsmara or Nguni bull). The frozen/thawed semen straws of Bonsmara and Nguni bulls were randomly selected and used under the same IVF conditions. The thawed bull’s sperm characteristics were examined by CASA before in vitro fertilization. Data were analysed using ANOVA. Treatment means were compared using the Fisher’s protected least significant difference t-test. There was no significant difference in oestrous response for the Bonsmara (83.0%) and Nguni (90.8%) cows, respectively. The Bonsmara cows recorded a significantly higher pregnancy rate (59.0%) compared with the Nguni (37.1%) cows (P < 0.05). Sperm traits such as total motility (TM), progressive motility and rapid were found to be positively correlated with conception rate (r = 0.06, 0.03, and 0.08, respectively; P < 0.01), although correlations were low. There was no difference in the average frozen–thawed sperm TM rate of Nguni (92.2%) and Bonsmara (81.0%). There was a lower fertilization rate following IVF with Bonsmara and Nguni bull sperm. In conclusion, Nguni cows had similar oestrous response as Bonsmara cows. The sperm traits from Bonsmara and Nguni bulls were found to be related to in vivo conception and in vitro fertilization rate when sperm cells were assessed by CASA technology. However, the pregnancy rate was lower in Nguni cows.

Fertilization and early embryonic development of in vitro matured metaphase I oocytes in patients with unexpected low oocyte maturity rate

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Nafiye Yılmaz ◽  
Şebnem Özyer ◽  
Derya Taş ◽  
Mehmet Caner Özer ◽  
Ayten Türkkanı ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Fertilization Rate ◽  
Early Embryonic Development ◽  
Oocyte Maturity ◽  
Significant Difference ◽  
Maturation Rate ◽  
Developmental Capacity ◽  
Metaphase I

Summary To determine the fertilization and embryonic potential of immature metaphase I (MI) oocytes in patients with low oocyte maturity rate in whom the percentage of mature oocytes obtained was less than 75% of the total retrieved ones. In vivo matured metaphase II (MII) oocytes (MII-ICSI, n = 244), and in vitro matured MI oocytes (MI-MII-ICSI, n = 202) underwent an intracytoplasmic sperm injection (ICSI) procedure. Maturation rate, fertilization rate and early embryonic development were compared in both groups. In total, 683 oocytes were collected from 117 ICSI cycles of 117 patients. Among them, 244 (35.7%) were mature MII and 259 (37.9%) were MI after the denudation process. Of those 259 MI oocytes, 202 (77.9%) progressed to MII oocytes after an incubation period of 18–24 h. The maturation rate was 77.9%. Fertilization rate was found to be significantly higher in the rescued in vitro matured MI oocyte group when compared with the in vivo matured MII oocyte group (41.6% vs 25.8%; P = 0.0006). However, no significant difference was observed in terms of cleavage rates on days 2 and 3 between the groups (P = 0.9126 and P = 0.5031, respectively). There may be unidentified in vivo factors on the oocyte maturation causing low developmental capacity in spite of high fertilization rates in the group of patients with low oocyte maturity rate. Furthermore, studies are needed to determine the appropriate culture characteristics as well as culture period and ICSI timing of these oocytes.

P–122 Benefit of use of theophilline in ICSI with testicular sperm

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Be. Khelif. Jerbi ◽  
I Chabchoub ◽  
S Sfaxi ◽  
M H Be. aribia ◽  
S Mnallah ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Sperm Motility ◽  
Time Interval ◽  
Sperm Selection ◽  
Testicular Sperm ◽  
Cumulative Pregnancy Rate ◽  
Significant Difference ◽  
Group A ◽  
Icsi Outcomes ◽  
Group B

Abstract Study question Would the use of theophylline have an effect on ICSI outcomes? Summary answer The cumulative pregnancy rate after the transfer of fresh and frozen embryos (FET) becomes more important justifying the addition of theophylline as an efficacy variable. What is known already Absolute immotile spermatozoa is one of the most important causes of reduced fertilization and pregnancy rates after ICSI, immotility of testicular spermatozoa is a physiological event resulting from metabolic sperm immaturity. Over the years, there have been numerous attempts to resolve this problem by identification of pharmacological agents that might improve sperm motility. In particular, theophylline turned out to be an effective tool in stimulating motility in human semen and identifying viable sperm in testicular sperm extraction sperm (TESE) patients. aim of this study is to evaluate ICSI outcomes after the use of theophylline to select viable spermatozoa Study design, size, duration This prospective, comparative randomised study was conducted in Fertillia ART center in Tunisia, between november 2017 to november 2020. All patients underwent ICSI cycles with testicular sperm were included. The exclusion criteria consist of cycles with female partner age &gt;42 years and/or cycles with no information about pregnancy outcomes. In our study, the cycles were categorized into two groups according to sperm selection method used in ICSI procdure. Participants/materials, setting, methods The present study include 678 cycles. This cohort was randomly divided into two groups according to method of spermatozoa selection. Treatements arms were performed by theophylline to improve sperm motility (Group A) and the Hyposmotic swelling (HOS) test to indicate sperm vitality (Group B). Clinical and biological parameters, the duration of sperm selection and ICSI outcomes were compared between the two groups. A statistical significant difference was accepted when the p value was &lt;0.05. Main results and the role of chance Baseline clinical parameters were found to be comparable in the two groups. No differences regarding number of oocytes retrieved or MII oocytes were reported . he sperm selection was easier in the group A (time interval : 12±7,2 minutes) than group B (time interval : 22±10 minutes) (p &lt; 0,05). No significative difference was observed between goups in the Fertilization rate (group A : 66% vs group B : 68%) ; Cleavage rate (group A : 76.20% vs. Group B : 75.56%) and Blasulation rate (group A : 63.60% vs. Group B 60.61%) p &gt; 0 ,05. The Cumulative Pregnancy rate for group A (31. 5%) was higher than group B (30%, p &gt; 0 ,05). Indeed, The rate of frozen cycles is significantly higher for group A compared to group B (30% vs. 4.5% p &lt; 0,05). Limitations, reasons for caution Need large sample size. Wider implications of the findings: Theophylline reduces significantly the time needed for sperm selection, Which positively affects the ICSI result. Trial registration number Not applicable

scholarly journals Q10 Co-enzyme Effect on Fetus Implantation in ART Cycles

2020 ◽  
Vol 8 (2) ◽  
pp. 17
Author(s):  
Aliyeh Ghasemzadeh ◽  
Samira Abdollahi Moghadam ◽  
Laya Farzadi ◽  
Mohammad Noori
Keyword(s):  
Pregnancy Rate ◽  
Coenzyme Q10 ◽  
Implantation Rate ◽  
Fertilization Rate ◽  
Small Sample ◽  
Control Group ◽  
Case Group ◽  
Significant Difference ◽  
Grade Ii

Background and Aim: Infertility is a common condition that affects many couples. Although current cutting-edge therapeutic methods have been found promising in this regard, their success still is not sufficiently high. Accordingly, researchers are working to develop new methods to augment the likelihood of successful results of Assisted Reproductive Technology (ART). Recently, coenzyme Q10 has been recognized an important and influential factor in the process of reproduction and some scarce studies have been along favorable results in this regard. This study aims to examine the effect of oral administration of coenzyme Q10 on embryo implantation in ART cycles. Methods and Materials: In this randomized, placebo-controlled clinical trial, a total of 128 infertile females who were candidates for ART were randomized in two groups receiving either a daily capsule of coenzyme Q10 by the commencement of gonadotropin through to pregnancy test result (case group, 64 patients), or placebo (control group, 64 patients). Finally, oocyte count, fertilization rate, embryo count, quality of embryos, transferred embryo count, implantation rate and pregnancy rate were compared between the two groups. Results: The mean age of the patients was 32.77±6.01 years (20-41) in the case group and 32.45±5.93 years (20-42) in the case group, with no significant difference between the two groups (p=0.77). In comparison between the case and control groups, in spite of better results in the case group, no significant difference was found in terms of oocyte count (10.47±7.16 and 9.38±7.52, respectively; p=0.40), fertilization rate (66.88±16.08 % and 66.73±21.50 %, respectively; p=0.96), embryo count (7.03±4.78 and 5.84±4.85, respectively; p=0.17), quality of embryos (71.9% grade I and 28.1% grade II in cases, 68.8% grade I and 31.3% grade II in controls; p=0.70) transferred embryo count (10.9% one, 56.3% two-three and 32.8% more than three in cases, 14% one, 56.3% two-three and 29.7% more than three in controls; p=0.84), implantation rate (8.39±12.73 % and 7.22±12.80 %, respectively; p=0.60), and pregnancy rate (34.4% and 26.6%, respectively; p=0.34). Conclusion: Although using coenzyme Q10, in comparison with placebo, was along with better results in terms of outcome variables of ART, the difference was statistically insignificant possibly because of a small sample size and short duration of the intervention.

scholarly journals The relationship between cannabis use and IVF outcome—a cohort study

2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Eden Har-Gil ◽  
Ayala Heled ◽  
Marjorie Dixon ◽  
Abdul Munaf Sultan Ahamed ◽  
Yaakov Bentov
Keyword(s):  
Cohort Study ◽  
Pregnancy Rate ◽  
Implantation Rate ◽  
Fertilization Rate ◽  
Cannabis Use ◽  
Ongoing Pregnancy ◽  
Female Reproduction ◽  
Ongoing Pregnancy Rate ◽  
Ivf Outcomes ◽  
Significant Difference

Abstract Background The effects of cannabis use on male and female reproduction have been the focus of scientific research for decades. Although initial studies raised concerns, more recent studies were reassuring. Considering the recent legalization of recreational use of cannabis in Canada, we sought to analyze IVF outcomes among users and non-users in a single IVF center. Methods This is a retrospective cohort study from a single IVF center assessing IVF outcomes among male-female, non-donor IVF patients that are either cannabis users or non-users. We analyzed the ongoing pregnancy rate as well as oocyte yield, fertilization rate, peak serum estradiol, sperm, and embryo quality. We used the Mann-Whitney test, chi-square test, and Kruskal-Wallis tests where appropriate. Results Overall, the study included 722 patients of which 68 (9.4%) were cannabis users, most defined as light users. The results of the study show similar implantation rate (40.74% vs. 41.13%) and ongoing pregnancy rate (35.2% vs. 29.1%) between the users and non-users, respectively. No significant difference between users and non-users in any of the other analyzed outcomes could be detected. Conclusions The results may provide some reassurance for the lack of any demonstrable detrimental effects of cannabis consumption on IVF outcomes. This study was limited by its retrospective nature, self-reporting of cannabis use, and a small user sample size. A larger prospective study is needed to validate its findings.

61 APPLICATION OF AN OPEN DEVICE TO VITRIFY EQUINE IN VITRO-PRODUCED EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 178 ◽  
Author(s):  
Y. H. Choi ◽  
I. C. Velez ◽  
B. Macías-García ◽  
K. Hinrichs
Keyword(s):  
Ethylene Glycol ◽  
Pregnancy Rate ◽  
Normal Pregnancy ◽  
Percentage Increase ◽  
Average Percentage ◽  
Base Medium ◽  
Significant Difference ◽  
Clinical Program

Recently, we reported a >50% normal pregnancy rate for vitrified/warmed equine expanded blastocysts after blastocoele collapse via micromanipulation and vitrification in fine-diameter (250-µm) pipettes (Choi et al. 2011 Theriogenology 76, 143). This vitrification system has also yielded high pregnancy rates after transfer of blastocysts produced by intracytoplasmic sperm injection and embryo culture (IVP embryos) in our clinical program (unpublished data). However, the pipettes are difficult to load and must be manipulated to expel the embryo after warming. To improve this technique, we modified the pipettes by cutting their tip lengthwise and flaming the surface smooth, and we investigated the use of these open devices (designated “Sujo”) for vitrification of equine IVP embryos. Embryos were held in 1.5 M ethylene glycol in DMEM/F-12 + 20% FBS for 5 min; then moved to 7 M ethylene glycol and 0.6 M galactose; and within 1 min loaded on a Sujo with a minimum amount of medium, plunged into liquid N2, and inserted into a 5-mL cryovial or 0.5-mL straw before being stored in liquid N2. Embryos were warmed by placing the Sujo tip in 0.3 M sucrose in base medium (Dulbecco’s phosphate buffered saline with 0.1% glucose, 36 mg L–1 of pyruvate, and 0.4% BSA). Embryos were left in this medium for 1 min and then moved to 0.15 M and then 0 M sucrose in base medium for 5 min each. In Experiment 1, we examined whether embryos stayed on the Sujo device. A total of 23 cleaved embryos were vitrified individually on Sujos; all of them were successfully recovered after warming. In Experiment 2, 18 IVP blastocysts were vitrified on Sujos (1 to 3 per Sujo), then warmed and cultured in 500-µL DMEM/F-12 + 20% FBS in an atmosphere of 5.5% CO2, 5% O2, and 89.5% N2 at 38.2°C for 3 days. All 18 embryos grew in vitro. The average percentage increase in diameter (mean ± SEM) for embryos vitrified at 1 per Sujo was 73% (from 167 ± 4 to 289 ± 16 µm, 11 embryos); 2 per Sujo, 98% (from 187 ± 27 to 387 ± 106 µm, 4 embryos); and 3 per Sujo, 96% (from 176 ± 4 to 342 ± 51 µm, 3 embryos). There was no significant difference in percentage growth among treatments (one-way ANOVA; SigmaPlot 11.0). In Experiment 3, 11 IVP embryos were vitrified after loading singly on Cryolocks®, then warmed and cultured as above. One embryo did not grow; the remaining 10 embryos grew an average of 73% (from 170 ± 3 to 294 ± 31 µm). In a preliminary study, 2 in vivo-recovered blastocysts (277 and 411 µm) were vitrified singly on Sujos and then warmed and cultured in vitro for 24 h under the above conditions increased in diameter to 594 and 729 µm, respectively. In conclusion, an open device can be used effectively for vitrification of equine IVP embryos. Further studies are needed to determine the efficiency of this system for vitrification of expanded equine blastocysts and the pregnancy rate of these embryos after transfer. Supported by the American Quarter Horse Foundation, the Link Equine Research Endowment Fund, Texas A&M University, and by Ms. Kit Knotts.

Fertilization following intracytoplasmic sperm injection of in vivo and in vitro matured oocytes from an Australian marsupial, the tammar wallaby (Macropus eugenii)

Zygote ◽  
2003 ◽  
Vol 11 (4) ◽  
pp. 339-346 ◽  
Author(s):  
Genevieve M. Magarey ◽  
Karen E. Mate
Keyword(s):  
Intracytoplasmic Sperm Injection ◽  
Germinal Vesicle ◽  
Tammar Wallaby ◽  
Fertilization Rate ◽  
Culture Conditions ◽  
Sham Injection ◽  
Significant Difference ◽  
Australian Marsupial

Conventional in vitro fertilization (IVF) techniques have been unable to produce normal embryos in any Australian marsupial, largely owing to problems with the early stages of sperm–oocyte binding. This study has used intracytoplasmic sperm injection (ICSI) of in vivo and in vitro matured tammar wallaby oocytes to bypass these processes and achieve fertilization in vitro. The fertilization rate (i.e. development to the two-pronuclei stage) of in vivo and in vitro matured oocytes following ICSI and sham injection was assessed at 17–19 h after injection. Fertilization occurred in 48% (45/93) of in vivo matured oocytes that were injected with spermatozoa. Significantly fewer sham-injected oocytes (6/82, P<0.005) and uninjected control oocytes (5/84, P<0.005) formed two pronuclei. In a direct comparison, the numbers of in vivo and in vitro matured oocytes that formed two pronuclei after ICSI were 22/28 (78.6%) and 23/40 (57.6%), respectively, which are not significantly different. There was also no significant difference in the nuclear response of in vivo and in vitro matured oocytes to sham injection. The numbers of oocytes forming a single pronucleus after sham injection were 10/24 (41.7%) and 24/37 (64.9) for in vivo and in vitro matured oocytes, respectively. Immature germinal-vesicle-stage oocytes were unable to decondense sperm injected during ICSI or to form pronuclei. These results demonstrate that both in vitro and in vivo matured tammar wallaby oocytes can be fertilized by ICSI. The success of ICSI not only offers the opportunity for fundamental analysis of marsupial fertilization but could, in conjunction with development of appropriate culture conditions and embryo transfer technologies, contribute to increased production of offspring from rare or valuable marsupials.

P–062 Annexin-V MACS method could be a good choice for sperm selection with high PLCZ1 expression and high blastocyst rate in high DFI male factor

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Saleh. Novin ◽  
Z Zandie ◽  
M Bakhtiari ◽  
R Aflatoonian
Keyword(s):  
Dna Fragmentation ◽  
Sperm Motility ◽  
Annexin V ◽  
Fertilization Rate ◽  
Good Choice ◽  
Semen Parameters ◽  
Sperm Selection ◽  
Male Factor ◽  
Significant Difference ◽  
Blastocyst Rate

Abstract Study question What effect does Annexin-V MACS method have on sperm selection by pLCZ1 expression and blastocyst rate in male DNA fragmentation? Summary answer AnnexinV-MACS method could be a good choice for sperm selection with high pLCZ1 expression and high blastocyst rate in high DNA fragmentation male factor. What is known already Sperm selection based on morphology and motility in ART techniques, is not enough for choosing the best sperm especially in male factor patients. In Annexin-V magnetic activated cell sorting (MACS) technique, apoptotic sperm are separated from non-apoptotic one by negative selection. So, this method can improved quality of compaction rate in embryo. PLCζ is oocyte activating factors that it starts oscillations of calcium ca2+ in oocyte and it has a significant effect on fertilization and implantation. Study design, size, duration Semen samples from 30 male factor infertile couples with high DFI (DFI&gt;30%) were selected and divided into two group (control and experimental) in each patient. Participants/materials, setting, methods Control was washed with DGC and experimental one was selected by MACS-DGC. Retrieved eggs in each patient, were divided in 2. Both group were injected by DGC and MACS respectively. Semen parameters and DFI (SCD test) were analyzed before and after processing. After ICSI, rate of fertilization, embryo development and blastocyst formation were evaluated. Real time PCR evaluate expression of PLCζ. Comparison between results of two groups was assessed by SPSS analysis. Main results and the role of chance Results showed that, sperm motility and morphology after MACS method (45%, 1.7%) was significantly higher than DGC method (40%, 1.1%) and before washing (35%, 0.9%). Percent of DFI in MACS group (36%) was significantly decreased compared to DGC (45%) and primitive group (55%). The number of oocytes were injected in DGC group was 93 and in MACS group was 111. Fertilization rate in both groups was almost the same (72.07% in MACS vs 73.11 in DGC). Rate of day 3 embryo with good grade in MACS group (72. 5%) was significantly higher than DGC (51.47%) (P &lt; 0.05). The blastocyst rate in MACS-DGC group (69.69%) was significantly increased compared to DGC group (48%). PLCζ gene expression in MACS-DGC was significantly higher than DGC group (p-value=0.046). Limitations, reasons for caution This experiment was performed invivo. Wider implications of the findings: Sperm selection by MACS-DGC method can improve sperm motility, morphology and reduce sperm DFI. No significant difference was observed in fertilization rate, but percent of high-quality embryo on days 3 and 5 was significantly higher by this method,also it can be suggested as a good choice for patients with highDF. Trial registration number Not applicable

Sign in / Sign up

Export Citation Format

Share Document