88 Evaluation of embryo transfer results using embryos cryopreserved in ethylene glycol for 8 years or in glycerol for 30 years

2019 ◽  
Vol 31 (1) ◽  
pp. 170
Author(s):  
C. Acevedo ◽  
S. Romo ◽  
C. López ◽  
A. Cortes-Mcnealy ◽  
M. I. Cruz-González ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Pregnancy Rate ◽  
Liquid Nitrogen ◽  
Embryo Transfer ◽  
Animal Health ◽  
Pregnancy Rates ◽  
Future Research ◽  
Embryo Viability ◽  
Exact Test ◽  
Significant Difference

Various permeating cryoprotectants, such as glycerol and ethylene glycol, have been used in the cryopreservation of embryos to help maintain cellular viability during indefinite and prolonged periods of storage in liquid nitrogen. The objective of this study was to compare the efficiency of glycerol (G) and ethylene glycol (EG) after storage in liquid nitrogen for a considerable period of time before transfer. The work was carried out in Palenque, Chiapas, Mexico. A total of 50 embryos were transferred, 24 Brahman (G) cryopreserved in the 1990s and 26 Brangus (EG) cryopreserved in 2010. Synchronous recipients were selected based on 3 characteristics: body condition (5-7, scale of 1-9), reproductive health, and multiparity. Recipient cows (n=62) were synchronized using a FTET protocol as follows. On Day 0, cows received a progesterone intravaginal device (CIDR) and 2mg of oestradiol benzoate IM. On day 8, the CIDR was removed and all cows received 25mg of dinoprost tromethamine (Lutalyse, Pfizer Animal Health, Montreal, Quebec, Canada), 200IU of eCG, and 0.5mg oestradiol cipionate IM. Day 10 was considered the day of oestrus and embryos were transferred (n=50) to the ipsilateral uterine horn of those recipients with a corpus luteum greater than 1.5cm in diameter on Day 17. The G embryos were produced with 4 bulls whereas the EG embryos were produced with 6 different bulls. The G straws were thawed for 12s in the air plus 12s in 20°C water. Embryos were immersed for 8min in a thawing solution containing 1.0M sucrose (ViGRO One-Step) and then transferred to holding medium (ViGRO Holding) for rehydration before loading into straws for embryo transfer. The EG embryos were thawed by allowing the straws to stand in air for 10s and then immersing them in a 30°C water bath for 10s and were transferred immediately. Pregnancy diagnosis 35 days after the transfer revealed 19 pregnancies of 50 embryos transferred (38%), distributed as 46% embryos in EG (12 pregnant of 26 transferred) and 29% embryos in G (7 pregnant of 24). A Fisher’s exact test was performed showing that no significant difference existed between groups (P>0.05). There was no effect of bull on pregnancy rates, and Brahman breed results by individual bull were 5 pregnancies of 13 (38%), 2 of 6 (33%), 0 of 4 (0%), and 0 of 1 (0%) for bulls I to IV, respectively. Pregnancy rate by Brangus bulls were 6 pregnancies of 7 (86%), 2 of 3 (67%), 2 of 4 (50%), 2 of 4 (50%), 0 of 4 (0%), and 0 of 3 (0%) for bulls 1 to 6, respectively. It is important to remember that the embryos cryopreserved in G remained in the nitrogen tank for more than 30 years, whereas the embryos cryopreserved in EG remained stored in liquid nitrogen for less than 10 years. Although pregnancy rate was numerically lower with Brahman embryos stored in G, pregnancy rates were considered acceptable considering the length of storage. Future research is needed with greater numbers and different breeds to determine whether G or EG will consistently produce higher embryo viability and pregnancies after storage for considerable periods before transfer.

scholarly journals Effect of different dosages of PG-600 on ovulation and pregnancy rates in ewes during the breeding season

2018 ◽  
Vol 3 (1) ◽  
pp. 429-432 ◽  
Author(s):  
Hayder Mohammed Hassan Habeeb ◽  
Timothy M Hazzard ◽  
Fred Stormshak ◽  
Michelle A Kutzler
Keyword(s):  
Pregnancy Rate ◽  
Breeding Season ◽  
Chorionic Gonadotropin ◽  
Repeated Measures ◽  
Animal Health ◽  
Pregnancy Rates ◽  
Analysis Of Covariance ◽  
Future Research ◽  
Nonbreeding Season ◽  
Estradiol 17Β

Abstract This study compared the reproductive effects of different dosages of PG-600 (Intervet/Merck Animal Health, Madison, NJ) during the breeding season of ewes. PG-600 is a single-dose injectable product labeled for estrous induction in swine, containing equine chorionic gonadotropin (80 IU/mL) and human chorionic gonadotropin (40 IU/mL). PG-600 is routinely used off-label for out-of-season estrous induction in sheep. However, at the most common dose administered to ewes (5 mL), PG-600 is likely to overstimulate the ovaries, resulting in reduced pregnancy rates. Following estrous synchronization with intravaginal progesterone and cloprostenol, Polypay ewes were treated with 5 mL PG-600 (T1; n = 8), 1.5 mL PG-600 (T2; n = 8), or 5 mL saline (C; n = 8) and then mated to rams. Jugular vein samples were collected prior to the PG-600 injection (0 hr) and at 2, 4, 8, 12, and 24 hr after injection. Serum estradiol-17β was determined by chemiluminescence and among groups using repeated measures analysis of covariance. Ovulation and pregnancy rates were determined by transrectal ultrasonography and compared by one-way ANOVA and chi-square, respectively. Estradiol-17β concentrations were greater in T1 compared to T2 and C (P < 0.001). Ovulation rate was greater (P < 0.001) but pregnancy rate was lower (P < 0.001) in the T1 compared to C and T2. These data confirm that a 5 mL dose of PG-600 administered to ewes during the breeding season overstimulates the ovaries, which may then reduce fertilization or embryo survival. Future research will focus on the effects of different dosages of PG-600 on pregnancy rate of ewes during the nonbreeding season.

216 THE RELATIONSHIP BETWEEN OXYGEN CONSUMPTION RATE AND PREGNANCY RATE OF BOVINE EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 225 ◽  
Author(s):  
N. Sakagami ◽  
K. Akiyama ◽  
Y. Nakazawa
Keyword(s):  
Oxygen Consumption ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Embryo Quality ◽  
Calf Serum ◽  
Pregnancy Rates ◽  
Bovine Embryos ◽  
Significant Difference ◽  
Consumption Rates ◽  
The Relationship

A precise evaluation of embryo quality is important to estimate the suitability of embryo transfer to recipient animal. Recently, an objective evaluation method was reported for bovine embryos, in which the oxygen consumption of embryos can be noninvasively determined by scanning electrochemical microscopy (SECM) (Shiku et al. 2001 Anal. Chem. 73, 3751–3758). Trimarchi et al. (2000 Biol. Reprod. 62, 1866–1874) suggested that the oxygen consumption reflects the cell number and mitochondrial activity of embryos. The objectives of this study were (1) to examine the oxygen consumption of in vivo-derived embryos by SECM, (2) to investigate the relationship between oxygen consumption and morphological estimation of embryos, and (3) to assess the correlation among the oxygen consumption, embryo viability, and pregnancy rates. Fifty-six embryos were collected from Japanese Black cattle, which were superovulated with a total dose of 20 mg porcine FSH (FSH-R; Kawasaki Pharmaceutical Co., Ltd., Tokyo, Japan) followed by AI. The qualities of collected embryos at the stage of compacted morulae (CM), early blastocysts (EB), and blastocysts (BL) on Day 7 after AI were categorized as grade 1 and grade 2, according to the IETS manual (2002). The oxygen consumption rates of embryos were evaluated by SECM, as previously described by Abe et al. (2004 J. Mamm. Ova Res. 21). Embryos were frozen by programmable freezer in Dulbecco's PBS containing 1.5 M ethylene glycol, 0.1 M trehalose, and 20% calf serum. They were thawed by holding the straws in air for 8 s and then immersing them in a 30°C water bath for 15 s. After thawing, the embryos were examined for oxygen consumption. Twenty-eight embryos were then cultured in TCM-199 supplemented with 20% fetal bovine serum and 0.1 mM β-mercaptoethanol for 24 h to assess the viability of embryos by re-expansion of blastocole. The remaining 28 embryos were transferred to recipients. The pregnancy rates were determined by rectal palpation on Day 70. Data were analyzed by ANOVA. The consumption rates of BL embryos on Day 7 were significantly higher (P < 0.05) than those of CM collected on the same day (0.84 vs. 1.29 × 10−14 mol s−1, respectively). A significant difference was also observed in consumption rates between grade 1 and 2 embryos at the BL stage (P < 0.05). After freezing–thawing, the average oxygen consumption rates of embryos were 0.52 × 10−14 mol s−1 for CM (n = 9), 0.67 × 10−14 mol s−1 for EB (n = 8), and 0.96 × 10−14 mol s−1 for BL (n = 11). The CM embryos with rates of < 0.5 × 10−14 mol s−1 and the EB and BL embryos with those < 0.6 × 10−14 mol s−1 did not show good morphological appearance after 24 h in culture. Pregnant animals were not obtained from embryos with rates <0.5 × 10−14 mol s−1 for CM (n = 5) and <0.7 × 10−14 mol s−1 for EB (n = 9). A high pregnancy rate (67%) was obtained from embryos with rates >1.0 × 10−14 mol s−1 for BL (n = 14). These results suggest that the measurement of oxygen consumption of embryos after embryo freezing and prior to embryo transfer may be useful for estimating embryo quality and suitability of embryo transfer.

37 EFFECT OF EMBRYO STAGE ON PREGNANCY RATE FOLLOWING DIRECT TRANSFER OF BOVINE EMBRYOS FROZEN IN ETHYLENE GLYCOL

2012 ◽  
Vol 24 (1) ◽  
pp. 131 ◽  
Author(s):  
J. F. Hasler
Keyword(s):  
Ethylene Glycol ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
The United States ◽  
Pregnancy Rates ◽  
Bovine Embryos ◽  
Embryo Freezing ◽  
Percentage Points ◽  
Embryo Stage

Annually, more than 400 000 in vivo-recovered bovine embryos are officially reported by members of the Canadian and American Embryo Transfer Associations. Between 65 and 70% of these embryos are cryopreserved and more than 95% are frozen in ethylene glycol (EG). Statistics on factors affecting embryo freezing are difficult to obtain because many cattle breeders/farmers no longer report pregnancy rates back to embryo transfer (ET) practitioners. Concerns are often expressed as to the optimal stage at which to freeze bovine in vivo-derived embryos. This is a retrospective analysis of results from 5 commercial ET programs (1 in the United States, 3 in Canada and 1 in the Netherlands) for which pregnancy data relative to embryo stage at freezing were made available. Embryos representing 4 stages of development, as defined by the IETS (4 = late morula, 5 = early blastocyst, 6 = mid blastocyst and 7 = expanded blastocyst) are included in the data. The number of embryos thawed and transferred ranged from 3954 to 24 827 for the 5 programs, with a total of 72 828. Embryos were frozen in either 1.5 M EG or 1.5 M EG + 0.1 M sucrose and exposure time to cryoprotectant before cooling ranged from 4 to 40 min. Pregnancy rates are shown in Table 1. Although the pregnancy rate for stage 6 embryos was only 2.6 and 3.2 percentage points lower than stages 4 and 5, respectively, these differences were highly significant and pregnancy rates for stage 6 embryos were lower than those for stages 4 and 5 in 4 of the 5 ET programs. The small decreased survival of stage 6 embryos is probably only moderately important in a commercial context. However, the pregnancy rate of stage 7 embryos was lower than all other stages for the combined dataset as well as in all 5 ET programs, with the difference between stages 5 and 7 ranging from 6.5 to 16.4 percentage points. Clearly, stage 7 embryos survive freezing at a significantly lower rate than stages 4, 5 and 6 and neither time of exposure to EG nor inclusion of sucrose in the freezing medium provided an obvious improvement. Although bovine ET practitioners routinely attempt to collect embryos on day 7 post-oestrus, recovery of stage 7 embryos cannot always be avoided. Further investigation into factors contributing to the decreased survival of stage 7 embryos is warranted. Table 1.Effect of embryo stage on pregnancy rate of bovine embryos frozen in EG

O-087 Embryos with higher mitochondrial DNA ratios show better clinical outcomes in single euploid embryo transfer

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
D Chen ◽  
C W Kao
Keyword(s):  
Mitochondrial Dna ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Implantation Rate ◽  
Single Embryo Transfer ◽  
Categorical Variables ◽  
Continuous Variables ◽  
Embryo Viability ◽  
Significant Difference ◽  
Euploid Embryo

Abstract Study question To assess whether there is a relationship between mitochondrial DNA content and implantation result. Summary answer The embryos with a higher mitochondrial DNA ratio increase pregnancy rate and implantation rate in single euploid embryo transfer. What is known already Mitochondria is an important organelle that generates energy during embryonic development. Recent literature points out that mitochondrial content and function may be related to implantation success and embryo viability. Some studies have linked increased ratios of mitochondrial DNA to aneuploidy, advanced maternal age, and euploid blastocyst with implantation failure, while others have failed to demonstrate similar findings. Study design, size, duration This study is a retrospective cohort study from 2016 to 2019, including 1465 single embryo transfer cycles. Participants/materials, setting, methods The involved embryos were biopsied on Day 5 or 6 and the mitochondrial DNA ratio of 1465 embryos was examined undergoing PGS/NGS. The mitochondrial DNA ratios were normalized for technical batch-to-batch variation. The mitochondrial DNA ratio between the implantation group and non-implantation group was statistically analyzed. Data were analyzed by the student’s t-test for continuous variables and Chi-square test for categorical variables. Main results and the role of chance The mitochondrial DNA ratio of embryos was no significant difference between different age spans ( p = 0.772) and ploidy (p = 0.224). D5 biopsied embryos, however, contained a significantly higher mitochondrial DNA ratio than D6 biopsied embryos (p < 0.0001). All of the single embryo transferred embryos were classified into two groups; implanted and non-implanted embryos. Results from 1465 transferred embryos show that the mitochondrial DNA ratio of implanted embryos was statistically significantly higher than non-implanted embryos (p = 0.0053). Besides, the cut-off values were established, dividing the transferred embryos into high and low mitochondrial DNA ratio groups. The pregnancy rate and implantation rate of the high mitochondrial DNA ratio group was higher than the low mitochondrial DNA ratio group: [Pregnancy rate] 74% vs. 63.5% (p = 0.0209); [Implantation rate] 57.3% vs. 50.8% (p = 0.1907). Limitations, reasons for caution The mitochondrial DNA ratios were analyzed by bioinformatics processing in Miseq reporter software (Illumina) files in the BAM and FASTQ format. Not sure if there is reproducibility in different sequencing platforms. Wider implications of the findings There still remains a lack of clarity regarding the relationship between mitochondrial function and transfer outcome. This retrospective study links an association between increased mtDNA content and increased implantation. Trial registration number not applicable

scholarly journals 70 RECONSTRUCTED BOVINE BLASTOCYSTS COMPRISING NUCLEAR TRANSFER-DERIVED INNER CELL MASS AND TROPHECTODERM FROM IVF EMBRYOS DO NOT IMPROVE IN VIVO DEVELOPMENT OF CLONES

2005 ◽  
Vol 17 (2) ◽  
pp. 185
Author(s):  
H.E. Troskie ◽  
F.C. Tucker ◽  
M.C. Berg ◽  
B. Oback ◽  
D.N. Wells ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Cell Line ◽  
Nuclear Transfer ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Pregnancy Rates ◽  
Reconstruction Procedure ◽  
Exact Test ◽  
Significant Difference

The cloning of cattle by somatic cell nuclear transfer (NT) is associated with a high incidence of abnormal placentation, excessive fluid accumulation in the fetal sacs (hydrops syndrome) and fetal overgrowth (Lee RSF et al. 2004 Biol. Reprod. 70, 1–11). Early embryonic loss in bovine NT pregnancies may also be due to immunological rejection (Hill JR et al. 2002 Biol. Reprod. 67, 55–63). As a means of overcoming placental abnormalities and improving pregnancy outcome in bovine NT, reconstructed blastocysts were produced by combining immunosurgically isolated inner cell masses (ICM) from Day 7 NT embryos with the trophectoderm (TE) of Day 7 IVF embryos. Oocytes for the production of NT and IVF embryos were obtained from abattoir-collected ovaries of dairy cows. The semen used for IVF was from the bull from which the cell line for NT was derived. The NT blastocysts were produced as described previously (Oback B et al. 2003 Cloning Stem Cells 5, 3–12) except that two one-cell embryos were aggregated together after NT (2NT). Blastocyst reconstruction was achieved using a modified procedure (Rorie RW et al. 1994 Vet. Record 135, 186–187). Embryos from four experimental groups were transferred individually to synchronized recipient heifers on Day 8 of culture: (1) ICM from 2NT embryos reconstructed with IVF TE (R-2NT, n = 15); (2) ICM from IVF embryos reconstructed with IVF TE (R-IVF, n = 15); (3) control 2NT (n = 10); and (4) control IVF (n = 10). Pregnancy rates were recorded and treatments compared using Fisher's exact test. After slaughter between Days 149 and 161 of gestation, morphometric measurements were determined for the fetuses, fetal organ weights, fluid volumes, and placentomes. Data were rank transformed; treatments were compared using Student's t-test with standard errors calculated from the pooled variation. Pregnancy rates on Day 35 were R-2NT (60%), R-IVF (47%), 2NT (90%), and IVF (10%). Pregnancy rates on Day 150 were R-2NT (40%), R-IVF (40%), 2NT (70%), and IVF (10%). The reason for the low IVF pregnancy rate was unknown. Previously, pregnancy rates using the same sire and cell line (but using Day 7 embryo transfer) on Day 35 were 63% (n = 40) and 69% (n = 42) for IVF and single, non-aggregated NT, respectively, and 50% and 33% for IVF and NT on Day 150. The single NT pregnancy rate was not significantly different from that for the 2NT embryos. There was no significant difference in pregnancy rates on Day 35 and Day 150 between R-2NT v. 2NT, R-2NT v. R-IVF, or 2NT v. R-IVF. The blastocyst reconstruction procedure did not have any impact on fetal development or influence pregnancy rates. All fetuses recovered were male. No significant differences were found between R-2NT and 2NT fetuses in terms of fetal weight, fluid volume, total placentome weight, and placentome numbers or in the relative and absolute weights of the brain, heart, liver, and kidneys. Thus, replacement of the TE in NT embryos with TE from IVF embryos did not overcome placental abnormalities or decrease fetal overgrowth prevalence.

170 POST-THAW VIABILITY OF IN VITRO-PRODUCED BOVINE EMBRYOS CULTURED EITHER IN HOST CAPRINE REPRODUCTIVE TRACTS OR IN VITRO

2006 ◽  
Vol 18 (2) ◽  
pp. 193
Author(s):  
S. Menges ◽  
C. Bormann ◽  
B. Stroud ◽  
D. Kraemer ◽  
M. Westhusin ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Pregnancy Rate ◽  
Reproductive Tract ◽  
Animal Health ◽  
Cumulus Cells ◽  
Bovine Embryos ◽  
Culture Methods ◽  
Vitro Fertilization ◽  
Significant Difference

In vitro culture of bovine embryos is usually associated with poor pregnancy rate following cryopreservation. The objective of this study was to compare the post-thaw viability of in vitro-produced bovine zygotes, cultured in vitro or in the reproductive tract of a host goat. Cumulus-oocyte complexes were matured in vitro, and in vitro fertilization was carried out with frozen-thawed semen as per standard laboratory procedures. At 18-20 h post-fertilization, zygotes were stripped of remaining cumulus cells and randomly separated into culture treatments. In three replicates, a total of 606 embryos were surgically transferred 12 to 24 h post-ovulation to the oviducts of an estrous-synchronized goat (VIVO) and 550 embryos were cultured in G1.3 for 72 h and then moved to G2.3 medium for 96 h and in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 (IVC). On Day 7, embryos were flushed from the excised tract with a 69.5% recovery rate or removed from culture. Embryos were classified according to IETS criteria with grades and stages recorded. All data were analyzed using the one-way analysis of variance and means were compared using Student's t-test. No differences were seen in the percentage of freezable quality embryos per total recovered between the two groups (34.3% vs. 32.3% for IVC and VIVO, respectively). However, there was a significant difference in the pre-freezing stage between the two culture groups (Stage 5.5 � 0.22 vs. Stage 4.8 � 0.26 for IVC and VIVO, respectively; P < 0.05), but no difference in the quality grade. All embryos greater than Stage 4, Grade 2 were frozen in groups of 5-10 in ethylene glycol with sucrose (Vigro Ethylene Glycol Freeze Plus; Bioniche Animal Health, Belleville, Ontario, Canada) in 0.25-mL straws. After thawing, embryo groups were washed, rehydrated, and incubated in G2.3 as above. Morphology was assessed by assigning grade and stage objectively at 24 h and 48 h post-thaw. Post-thaw viability in vitro was not different between groups (73.4% vs. 72.7% for IVC and VIVO, respectively). The average changes in morphology post-thaw from pre-freezing to 24 h and from 24 h to 48 h within each freezing group were determined. There was no significant difference in the mean change in stage (0.67 � 0.15 vs. 0.82 � 0.17 at 24 h and 0.31 � 0.09 vs. 0.37 � 0.10 at 48 h for IVC and VIVO, respectively) or grade (0.60 � 0.15 vs. 0.41 � 0.17 at 24 h and 0.03 � 0.06 vs. 0.14 � 0.07 at 48 h for IVC and VIVO, respectively) at either observation point. These results suggest that culture of in vitro-fertilized bovine embryos in the caprine reproductive tract did not alter post-thaw development or improve post thaw viability compared to in vitro cultured controls. However, morphological evaluation is too subjective to successfully predict pregnancy rate after transfer; therefore, further study is needed to determine if there are differences in pregnancy rates between these culture methods.

106 PREGNANCY RATE AFTER EMBRYO TRANSFER OF IN VIVO-PRODUCED OVINE EMBRYOS CRYOPRESERVED BY SLOW FREEZING OR VITRIFICATION

2010 ◽  
Vol 22 (1) ◽  
pp. 212
Author(s):  
N. Mucci ◽  
F. Hozbor ◽  
G. G. Kaiser ◽  
E. Sanchez ◽  
R. H. Alberio
Keyword(s):  
Ethylene Glycol ◽  
Pregnancy Rate ◽  
Liquid Nitrogen ◽  
Embryo Transfer ◽  
Slow Freezing ◽  
Embryo Cryopreservation ◽  
Chi Square ◽  
Chi Square Test

Although slow freezing is the method of choice to cryopreserve in vivo-produced ovine embryos, vitrification has became an alternative procedure mostly developed for in vitro-produced bovine embryos. The aim of this work was to compare pregnancy rates after cryopreservation of in vivo-produced ovine embryos with slow freezing or open pulled straw (OPS) vitrification method. Ewes were synchronized using intravaginal sponges containing 60 mg of medroxyprogesterone acetate for 14 d. Superovulation was performed using a total dose of 176 IU of ovine FSH (Ovagen), in 6 decreasing doses (i.m.) from Day 12 to 14 of treatment (Day 0 = sponge placing). Ewes were hand mated with 2 rams of proven fertility. Embryos were recovered 6 days after estrous detection by surgical procedure, evaluated under stereomicroscope, and randomly assigned to the cryopreservation treatments. Slow freezing was performed in D-PBS supplemented with 1.78 M ethylene glycol, 0.1 M sucrose, 4 mg mL-1 of BSA, and 20% serum. Embryos were loaded into 0.25-mL plastic straws and placed into a -7°C methanol bath chamber. After seeding embryos were cooled to -35°C at a rate of 0.5°C/min and then stored in liquid nitrogen. Thawing was performed by placing the straws in a 30°C water bath for 30 sec. Vitrification was performed by using the OPS method (Vajta et al. 1998) with minor modifications. Embryos were incubated in D-PBS supplemented with 1.78 M ethylene glycol, 1.3 M DMSO for 3 min and then transferred for 25 s in vitrification solution of D-PBS with 3.56 M ethylene glycol, 2.6 M DMSO, and 0.5 M sucrose, loaded in a 1 mL drop in the OPS, and immediately submerged into and stored in liquid nitrogen. Warming was performed in D-PBS plus 0.25 M sucrose for 5 min and then into D-PBS plus 0.15 M sucrose for another 5 min. Before embryo transfer, the presence of corpus luteum (CL) was detected by laparoscopic examination. One embryo per recipient was surgically transferred in the apical extreme of the uterine horn ipsilateral to the CL. Pregnancies were determined by ultrasonography 41 days after embryo transfer. Data were analyzed using the chi-square test. We found 47.8% pregnancy rate using slow freezing (11/23) and 43.5% pregnancy rate using OPS vitrification (10/23). Statistical differences were not detected (P = 0.09). We conclude that vitrification by OPS system, with minor modifications, is a suitable procedure for in vivo-produced ovine embryo cryopreservation.

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 &lt; 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 &lt; 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

Human oocyte vitrification with corona radiata, in autologous follicular fluid supplemented with ethylene glycol, preserves conventional IVF potential: birth of four healthy babies

2014 ◽  
Vol 26 (7) ◽  
pp. 1001 ◽  
Author(s):  
Xian-Hong Tong ◽  
Li-Min Wu ◽  
Ren-Tao Jin ◽  
Hong-Bing Luan ◽  
Yu-Sheng Liu
Keyword(s):  
Ethylene Glycol ◽  
Embryo Transfer ◽  
Follicular Fluid ◽  
Control Group ◽  
Human Oocyte ◽  
Oocyte Vitrification ◽  
Human Oocytes ◽  
Corona Radiata ◽  
Fertilisation Rate ◽  
Significant Difference

The aim of this study was to examine the effects of vitrification with autologous follicular fluid (AFF) supplemented with ethylene glycol (EG) and sucrose on human oocytes with corona radiata. A total of 182 human oocytes with corona radiata from fifteen infertile patients were vitrified using either equilibration solutions (ES) and vitrification solution (VS) consisting of AFF, EG and sucrose (AFF group, n = 67) or commercial ES and VS (control group, n = 115). All oocytes were thawed in the next cycle, with surviving oocytes being inseminated by conventional IVF. The clinical outcome of vitrified–warmed oocytes by both vitrification methods was analysed retrospectively. In the AFF group, six patients received embryo transfer, with three couples taking four healthy babies home. In the control group, nine patients received embryo transfer, with four couples taking five healthy babies home. There was no significant difference in the survival rate (91.0 vs 92.2%), two pronuclei (2PN) fertilisation rate (73.8 vs 73.6%), cleavage rate (100 vs 100%), top-quality embryo rate (62.2 vs 59.2%), clinical pregnancy rate (50.0 vs 44.4%), implantation rate (33.3 vs 25%) or take-home baby rate (50.0 vs 44.4%) between the AFF group and the control group, respectively. These results show that AFF supplemented with EG and sucrose is an efficient, cost-effective cryoprotectant for human oocyte cryopreservation. A corona radiata on vitrified–warmed oocytes retains the oocytes’ fertilisation capability in conventional IVF.

scholarly journals Comparative Study Between Mock Embryo Transfer Prior To The Treatment Cycle and Real Embryo Transfer In In Vitro Fertilization

KnE Medicine ◽  
2016 ◽  
Vol 1 (1) ◽  
Author(s):  
Hilma Putri Lubis
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Treatment Cycle ◽  
Uterine Cavity ◽  
Pregnancy Rates ◽  
Clinical Pregnancy ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Clinical Pregnancy Rates

<p><strong>Introduction</strong><strong></strong></p><p>A trial or mock embryo transfer (ET) may influence pregnancy rates and it performed prior to ET allows the clinician to assess the uterine cavity and the utero-cervical angle. The aim of this study is to compare the consistency of the type of ET in mock ET with real ET.</p><p><strong>Material &amp; Methods</strong></p><p>A retrospective comparative analysis of  patients who underwent in vitro fertilization or ICSI cycle from January 2014 to December 2014 in Halim Fertility Center was done. The type of transfer was divided into two groups: ‘easy’ or ‘difficult’. An easy ET was defined as a transfer that occurred without the use of manipulation or other instrumentation and difficult ET was considered when additional instrumentation was required.</p><p><strong>Results</strong></p><p>From the study, 103 patients who underwent Mock-ET, we  found 58 patients (56.3%) with easy ET and 45 patients (43.7%) with difficult ET, which with hard catheter ET in 17 patients (16.5%), with osfander assistance in 20 patients (19.4%) and with stylet in 8 patients (7,8%). 58 patients with Easy Mock ET group were entirely easy real ET (100%) and 45 patients with difficult Mock ET group also entirely were difficult real ET (100%). The Statistical analysis shows no significant difference between the mock ET and real ET groups (p&gt;0,05). In easy real ET, clinical pregnancy rates were 32.8% and in difficult real ET, clinical pregnancy rates were 26.7% with no significant difference between the  groups (p&gt;0,05).</p><p><strong>Conclusion:</strong></p><p>Mock ET prior to the treatment cycle is consistent with real ET.</p>

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