Endometrial microbiome in women with multiple failed in vitro fertilization cycles

2021 ◽  
Vol 20 (3) ◽  
pp. 5-11
Author(s):  
V.V. Barinova ◽  
◽  
N.B. Kuznetsova ◽  
I.O. Bushtyreva ◽  
K.M. Sokolova ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Gardnerella Vaginalis ◽  
Comamonas Testosteroni ◽  
Bifidobacterium Adolescentis ◽  
Vitro Fertilization ◽  
Healthy Women ◽  
High Concentrations ◽  
Group 2 ◽  
Group 1

Objective. To assess endometrial microbiome in women with infertility and multiple failed in vitro fertilization (IVF) cycles. Patients and methods. The study included 42 women; group 1 consisted of 22 women aged 20 to 42 with infertility and repeated unsuccessful IVF cycles; in group 2 (control), there were 20 healthy women aged 20 to 42 years, planning pregnancy. Microbiome samples for the study were taken from 20 to 24 days of the menstrual cycle. Results. Higher relative concentrations of Lactobacillus iners, Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillus crispatus, Prevotella melaninogenica, Bacteroides vulgatus, Corynebacterium bouchesdurhonense, Bacteroides caccae, Bifidobacterium gallinarum, Bifidobacterium adolescentis were revealed in the group of healthy women without aggravating factors in obstetric and gynecological medical history. Higher relative concentrations of Methylobacterium aerolatum and Comamonas testosteroni were found in women in group 1. A distinctive feature of endometrial microbiota in women in this group was the presence of Streptococcus spp. and Gardnerella vaginalis in low concentrations. The average relative representation of the genus Lactobacillus was 34.4% in group 1 and 63.0% in group 2. In general, the composition of the uterine microbiome contained bacteria characteristic of oral and intestinal biotopes. Conclusion. Women with multiple failed IVF cycles have greater biodiversity than healthy women. The presence of high concentrations of Lactobacillus may be a marker of favorable reproductive outcomes. Key words: endometrial microbiome, failure, in vitro fertilization, Lactobacillus

scholarly journals Effect of Cumulus cell co-culture and Protein Supplement on Success of in vitro Fertilization and Development of Pre-implanted Embryos in mice

2005 ◽  
Vol 10 ◽  
pp. 31
Author(s):  
Muhammad-Baqir M-R. Fakhrildin
Keyword(s):  
In Vitro Fertilization ◽  
Embryonic Development ◽  
Amniotic Fluid ◽  
Cumulus Cells ◽  
Protein Source ◽  
Vitro Fertilization ◽  
Group 2 ◽  
Normal Embryonic Development ◽  
Group 1

Successful oocyte fertilization and normal embryonic development of mice were considered the most important diagnostic criteria for the safety of materials and tools used for human in vitro fertilization and embryo transfer (IVF-ET). Therefore, we studied the influence of cumulus cells co-culture and protein supplement within culture medium on percentages of in vitro fertilization (IVF) and normal development of early stages of mouse embryo later. Oocytes were collected and treated with hyaluronidase to remove cumulus cells. Oocytes were divided into four groups namely: Group-1: Oocytes incubated within modified Earl’s medium (MEM) supplied with 10% inactivated bovine amniotic fluid as a protein source and cumulus cells; Group-2: Oocytes incubated with MEM supplied with cumulus cells only; Group-3: Oocytes incubated with MEM supplied with 10% inactivated bovine amniotic fluid only; and Group-4: Oocytes  incubated with MEM free of both protein source and cumulus cells. For IVF, 5-6 oocytes were incubated with active spermatozoa under paraffin oil for 18-20 hours at 37° oC in 5% CO2. Percentages of IVF and embryonic development were then recorded. Best results for IVF and normal embryonic development were achieved from oocytes of Group-1 when compared to the other groups. As compared to Group-1, the percentage of IVF for Group-2 and Group-3 were decreased insignificantly and significantly (P<0.002), respectively. Significant (P<0.01) reduction in the percentages of IVF and normal embryonic development were reported in Group-4 as compared to Group-1. Therefore, it was concluded that the presence of cumulus cells co-culture and bovine amniotic fluid as a protein source within culture medium may have an important role on the fertilizing capacity of spermatozoa and oocytes and normal development of pre-implanted mouse embryo later.  

scholarly journals Re-evaluation of the value of sperm morphology in classical in vitro fertilization in a Northeastern Chinese population

2019 ◽  
Vol 47 (9) ◽  
pp. 4134-4142 ◽  
Author(s):  
Dong-liang Zhu ◽  
Hong-guo Zhang ◽  
Rui-xue Wang ◽  
Yu-ting Jiang ◽  
Rui-zhi Liu
Keyword(s):  
In Vitro Fertilization ◽  
Chinese Population ◽  
Sperm Morphology ◽  
Semen Analysis ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Group 2 ◽  
Normal Fertilization ◽  
Group 1

Objective This study aimed to re-evaluate the clinical value of a 4% cut-off threshold of sperm morphology in in vitro fertilization (IVF) in a cohort of a Northeastern Chinese population. Methods A total of 375 IVF cycles that met strict inclusion criteria were included. These cycles were conducted with semen analysis and oocyte fertilization. A total of 188 embryo-transferred cycles proceeded. According to sperm morphology, 375 cycles were divided into group 1 (329 cycles, <4% normal sperm morphology rate [NSMR]) and group 2 (46 cycles, ≥4% NSMR), and 188 transferred cycles into group A (151 cycles, < 4% NSMR) and group B (37 cycles, ≥4% NSMR). Results The fertilization and normal fertilization rates were significantly lower in group 1 than in group 2. The normal fertilization rate was significantly correlated with an NSMR < 4% or ≥4%, but the fertilization rate was not significantly correlated with the NSMR. No significant differences were found in pregnancy outcomes between groups A and B. Conclusions This study suggests that infertile patients with an NSMR < 4% are more likely to have a poor normal fertilization status in IVF.

scholarly journals A Flexible Multidose GnRH Antagonist versus a Microdose Flare-Up GnRH Agonist Combined with a Flexible Multidose GnRH Antagonist Protocol in Poor Responders to IVF

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Gayem İnayet Turgay Çelik ◽  
Havva Kömür Sütçü ◽  
Yaşam Kemal Akpak ◽  
Münire Erman Akar
Keyword(s):  
Gnrh Agonist ◽  
Gnrh Antagonist ◽  
Poor Responders ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Gnrh Antagonist Protocol ◽  
Group 2 ◽  
Antagonist Protocol ◽  
Group 1

Objective. To compare the effectiveness of a flexible multidose gonadotropin-releasing hormone (GnRH) antagonist against the effectiveness of a microdose flare-up GnRH agonist combined with a flexible multidose GnRH antagonist protocol in poor responders to in vitro fertilization (IVF).Study Design. A retrospective study in Akdeniz University, Faculty of Medicine, Department of Obstetrics and Gynecology, IVF Center, for 131 poor responders in the intracytoplasmic sperm injection-embryo transfer (ICSI-ET) program between January 2006 and November 2012. The groups were compared to the patients’ characteristics, controlled ovarian stimulation (COH) results, and laboratory results.Results. Combination protocol was applied to 46 patients (group 1), and a single protocol was applied to 85 patients (group 2). In group 1, the duration of the treatment was longer and the dose of FSH was higher. The cycle cancellation rate was significantly higher in group 2 (26.1% versus 38.8%). A significant difference was not observed with respect to the number and quality of oocytes and embryos or to the number of embryos transferred. There were no statistically significant differences in the hCG positivity (9.5% versus 9.4%) or the clinical pregnancy rates (7.1% versus 10.6%).Conclusion. The combination protocol does not provide additional efficacy.

scholarly journals Association Between Serum Oestradiol Level on the Human Chorionic Gonadotrophin Administration Day and Neonatal Birthweight After in Vitro Fertilization-embryo Transfer: A Retrospective Study of 3659 Singleton Live Births

2020 ◽  
Author(s):  
Yu Liu ◽  
Jing Li ◽  
Yihong Guo
Keyword(s):  
Retrospective Study ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Vitro Fertilization ◽  
Group 4 ◽  
Group 3 ◽  
Group 2 ◽  
Ivf Et ◽  
Group 1

Abstract BackgroundOestradiol, an important hormone in follicular development and endometrial receptivity, is closely related to clinical outcomes of fresh in vitro fertilization-embryo transfer (IVF-ET) cycles. A supraphysiologic E2 level is inevitable during controlled ovarian hyper-stimulation (COH), and its effect on the outcome of IVF-ET is controversial. The aim of this retrospective study is to evaluate the association between elevated serum oestradiol (E2) levels on the day of human chorionic gonadotrophin (hCG) administration and neonatal birthweight after IVF-ET cycles.MethodsThe data of 3659 infertile patients with fresh IVF-ET cycles were analysed retrospectively between August 2009 and February 2017 in First Hospital of Zhengzhou University. Patients were categorized by serum E2 levels on the day of hCG administration into six groups: group 1 (serum E2 levels≤1000 pg/mL, n=230), group 2 (serum E2 levels between 1001 and 2000 pg/mL, n=524), group 3 (serum E2 levels between 2001 and 3000 pg/mL, n=783), group 4 (serum E2 levels between 3001 and 4000 pg/mL, n = 721), group 5 (serum E2 levels between 4001 and 5000 pg/mL, n=548 ), and group 6 (serum E2 levels > 5000 pg/mL, n=852). Univariate linear regression was used to evaluate the independent correlation between each factor and outcome index. Multiple logistic regression was used to adjust for confounding factors.ResultsThe LBW rates were as follows: 3.0% (group 1), 2.9% (group 2), 1.9% (group 3), 2.9% (group 4), and 2.0% (group 6) (P =0.629), respectively. There were no statistically significant differences in the incidences of neonatal LBW among the six groups. We did not detect an association between peak serum E2 level during ovarian stimulation and neonatal birthweight after IVF-ET.ConclusionThe results of this retrospective cohort study showed that serum E2 peak levels during ovarian stimulation were not associated with birth weight during IVF cycles. In addition, no association was found between higher E2 levels and increased LBW risk. Our observations suggest that the hyper-oestrogenic milieu during COS does not seem to have adverse effects on the birthweight of offspring after IVF.

scholarly journals 253 COMPARISON OF THREE DIFFERENT IVF PROTOCOLS FOR BOVINE OOCYTES

2005 ◽  
Vol 17 (2) ◽  
pp. 277
Author(s):  
S. Romo ◽  
J. Pryor ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney
Keyword(s):  
Embryo Culture ◽  
Culture Media ◽  
Sperm Concentration ◽  
Vitro Fertilization ◽  
Group 3 ◽  
Group 2 ◽  
The Given ◽  
Sperm Separation ◽  
Group 1

Recently, the development of commercially available defined media and sperm centrifugation gradients has offered new possibilities for increasing the efficiency of commercial in vitro fertilization (IVF) systems. The objective of this study was to compare three different IVF protocols using two different separation gradients, two fertilization media, and two embryo culture media, as follows: Group 1. sperm separation (SS): Percoll (Sigma, St. Louis, MO, USA), fertilization medium (FM): TALP-Fert (TFM), embryo culture media (ECM): G1/G2 (version 3, Vitrolife, Englewood, CO, USA). Group 2. SS: Percoll, FM: Bovine vitro Fert (Cook, Brisbane, Australia), ECM: Bovine vitro Blast/Bovine vitro Cleave (Cook); and Group 3. SS: EquiPure (Nidacon, Spectrum Technologies, Healdsburg, CA, USA), FM: TFM, ECM: G1/G2. Oocytes were obtained from slaughterhouse ovaries and matured in vitro (Looney et al. 1994 Theriogenology 41, 67). IVF was conducted using frozen/thawed semen from one bull. Semen was separated by centrifugation at 700g for 30 min in the given density gradients; Percoll was used in a 45% to 90% gradient. Sperm viability after separation was assessed by fast-green/eosin stain (Sigma). IVF was carried out in 0.5 mL of the given fertilization medium supplemented with PHE1 and heparin (10 μg/mL), in humidified 5% CO2 in air atmosphere at 38.7°C. Final sperm concentration in the IVF wells was 1 × 106/mL. In Experiment 1, a total of 368 oocytes (2 replicates) were fixed and stained (Hoechst 33342, Sigma) 24 h post-IVF to assess sperm penetration (Group 1, n = 128, Group 2, n = 108, Group 3, n = 132). In Experiment 2, a total of 400 embryos (2 replicates) were cultured in 0.5 mL of the given culture medium under mineral oil in a 5% O2, 5% CO2, 90% N2 atmosphere at 38.7°C with high humidity for 112 h before fixation and staining. Embryos in Groups 1 (n = 129) and 3 (n = 139) and Group 2 (n = 132) were changed to G2 and Cleave media, respectively, at 84 h. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 218 vs. 383 × 106 for EquiPure; P < 0.05), but resulted in higher total motility (60% vs. 41%, respectively; P < 0.05) and higher viability (93% vs. 70%, respectively; P < 0.05) of separated sperm. In Experiment 1, rates of normal fertilization were significantly lower for Group 3 (58%) than for Groups 1 and 2 (74% and 77%, respectively, P < 0.05). In Experiment 2, rates of development to <8, 9 to 16, and >16 cells at 112 h were not significantly different among groups (43, 48, and 46% for Group 1; 22, 18, and 31% for Group 2; and 35, 34, and 23% for Group 3, respectively; P > 0.1). These results indicate that the commercial separation medium, EquiPure, may be associated with lowered sperm motility, viability, and fertilization rates when compared to a standard medium (Percoll) for bovine sperm separation. Commercial fertilization and embryo culture media (Bovine vitro Fert, Cleave, and Blast) provided equivalent embryo development to that currently in use by our laboratory (TFM, G1/G2).

313 APOPTOSIS IN PARTHENOGENETIC PIG EMBRYOS PRODUCED BY DIFFERENT METHODS

2006 ◽  
Vol 18 (2) ◽  
pp. 264
Author(s):  
J. Hyslop ◽  
Z. Machaty
Keyword(s):  
In Vitro Fertilization ◽  
Cell Number ◽  
Apoptotic Cells ◽  
Total Cell Number ◽  
Vitro Fertilization ◽  
Tunel Reaction ◽  
Group 3 ◽  
Blastocyst Rate ◽  
Group 2

Apoptosis or programmed cell death is a process during which cells die in a controlled fashion in response to a variety of stimuli. Apoptosis has been demonstrated to occur during pre-implantation development both in vivo and in vitro and it is believed to contribute to early embryonic loss. It is also believed that parthenogenetic embryos generally have a lower developmental potential compared to those derived from fertilization. In the present study we investigated the rate of apoptosis in parthenogenetic pig embryos produced by activating oocytes through various methods. Pig oocytes were collected from slaughterhouse ovaries and matured in vitro. Parthenogenetic development was induced by three different methods. In Group 1, oocytes were activated by two consecutive electrical pulses. In Group 2, oocytes were electroporated and then incubated for 4 h in 5 mM butyrolactone I, a specific inhibitor of cyclin dependent kinases. In Group 3, electroporated oocytes were incubated for 5 h in cycloheximide, a protein synthesis inhibitor. Activated oocytes were cultured for 7 days in NCSU-23 medium. At the end of the culture period the embryos were fixed in 4% paraformaldehyde and permeabilized in 0.1% Triton X-100 with sodium citrate. They were then incubated in a TUNEL reaction medium that specifically identifies apoptotic nuclei by labeling fragmented DNA with a fluorochrome. Blastocysts produced by in vitro fertilization and DNase I-treated embryos were used as controls. The proportions of apoptotic cells were compared using ANOVA. Forty-three blastocysts were analyzed for apoptotic activity in the electroporation group. These embryos had a blastocyst rate of 33.6 ± 8.7%, total cell number of 31.9 ± 13.2, and an average of 2.7 ± 2.2 apoptotic cells per embryo; the rate of apoptosis was 9.1 ± 7.1%. Twenty-eight blastocysts were used for the TUNEL reaction in the group where activation was induced with the combined treatment of electroporation and butyrolactone (Group 2). On average, the blastocyst rate was 54.5 ± 6.4% and blastocysts contained 27.4 ± 9.6 cells of which 2.8 ± 3.9 were apoptotic; the percentage of apoptosis for this group was 10.0 ± 12.1%. In the cycloheximide treated group (Group 3), the onset of apoptosis was investigated using 29 blastocysts. The blastocyst rate was 38.2 ± 15.9% with an average total cell number of 27.2 ± 11.4%. The TUNEL assay revealed that the mean number of apoptotic cells per embryo in these blastocysts was 2.1 ± 1.5, representing 9.0 ± 7.6% apoptotic cells. The blastocysts (n = 14) produced by in vitro fertilization had a blastocyst rate of 18.0 ± 5.1% and 29.9 ± 12.0 cells; 9.2 ± 8.1% of these cells (2.6 ± 2.2 cells per embryo) showed signs of apoptosis. All nuclei in the DNase I-treated embryos showed positive signals following the TUNEL reaction. The results confirm previous findings that apoptosis occurs in blastocyst stage embryos. There was no difference in the percentage of apoptotic cells between embryos whose development was triggered by different oocyte activation methods; the rate of apoptosis in parthenogenetic blastocysts was similar to that in blastocysts produced by in vitro fertilization.

scholarly journals Efecto De La Estimulación Endometrial En La Tasa De Exito En Pacientes Sometidas A Fertilización In Vitro [Effect Of Endometrial Stimulation On Success Rate In Patients Submitted To In Vitro Fertilization ]

2020 ◽  
Vol 24 (2) ◽  
Author(s):  
Lorna M Frazer Moreira ◽  
Felipe A Morales Martinez ◽  
María E. Monrreal Alanís ◽  
Otto H. Valdés Martinez ◽  
Cesar D. Castro Reyes ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Success Rate ◽  
Endometrial Biopsy ◽  
Clinical Pregnancy ◽  
Vitro Fertilization ◽  
The Third ◽  
Endometrial Injury ◽  
Group 2 ◽  
Vitro Effect

<h1><em>Resumen </em></h1><p><em>Introducción</em>: A pesar de los avances en las técnicas de reproducción asistida, la tasa de éxito en fertilización in vitro (FIV) sigue siendo modesta. La lesión endometrial, mediante histeroscopía o biopsia, previo a un ciclo de FIV, ha sido propuesta como una técnica que aumenta la probabilidad de implantación exitosa y embarazo en mujeres sometidas a FIV. <em>Objetivo</em>: Estudiar el efecto de la estimulación endometrial en la tasa de éxito de FIV. <em>Métodos</em>: Se realizó un estudio unicéntrico observacional, en pacientes sometidas a su primer ciclo de FIV. Se incluyeron mujeres entre las edades de 18-40 años que planeaban someterse a FIV con sus propios ovocitos sin exposición reciente a procedimientos disruptivos uterinos. Las pacientes elegibles se dividieron en 3 grupos. El primer grupo incluyó las que se sometieron a una histeroscopía diagnóstica previa a su ciclo de FIV, el segundo grupo incluyó aquellas que se sometieron a biopsia endometrial con cánula de Pipelle previo al ciclo de FIV y el tercer grupo incluyó a las pacientes sin intervención endometrial. <em>Resultados</em>: Se estudiaron un total de 68 pacientes (10 en el grupo 1, 27 en el grupo 2 y 31 en el grupo 3). La tasa de embarazo clínico fue estadísticamente similar en los tres grupos (33.3% para el primer grupo, 15.7% para el segundo y 16.6% para el tercero, p = 0.93). <em>Conclusión</em>: La estimulación endometrial antes del primer ciclo de FIV no ofreció beneficios adicionales en relación con implantación exitosa y/o tasas clínicas de embarazo. </p><p align="left">Palabras clave: lesión endometrial, fertilización in vitro, implantación, infertilidad, nacido vivo</p><p align="left"> </p><h1>Abstract</h1><p><em>Background</em>: Despite advances in assisted reproduction techniques, the in vitro fertilization (IVF) success rate still remains modest. Endometrial injury, through hysteroscopy or biopsy, prior to an IVF cycle, has been proposed as a technique that increases the likelihood of successful implantation and pregnancy in women undergoing IVF. <em>Aim and Objective</em>: To study the effect of endometrial stimulation on the success rate of IVF. <em>Methods</em>: An ambispective observational unicentric study was conducted in patients undergoing their first IVF cycle. Women between the ages of 18-40 who planned to undergo IVF with their own oocytes without recent exposure to uterine disruptive procedures were included. Eligible patients were divided into 3 groups. The first group included those who underwent a diagnostic hysteroscopy prior to their IVF cycle, the second group included those who underwent endometrial biopsy with a Pipelle cannula prior to the IVF cycle and the third group included patients without endometrial intervention. <em>Results</em>: A total of 68 patients were studied (10 in group 1, 27 in group 2 and 31 in group 3). The clinical pregnancy rate was statistically similar in the three groups (33.3% for the first group, 15.7% for the second group and 16.6% for the third group, p = 0.93). <em>Conclusion</em>: Endometrial stimulation before the first IVF cycle did not offer additional benefits in relation to successful implantation and / or clinical pregnancy rates</p><p align="left">Keywords: endometrial injury, in vitro fertilization, implantation, infertility, live birth</p>*****

308 EFFECT OF GONADOTROPIN TREATMENT ON OOCYTE COLLECTION AND IN VITRO FERTILIZATION IN THE BAN MINIPIG

2007 ◽  
Vol 19 (1) ◽  
pp. 269 ◽  
Author(s):  
B. X. Nguyen ◽  
T. Nagai ◽  
K. Kukuchi ◽  
N. T. Uoc ◽  
M. Ozawa ◽  
...  
Keyword(s):  
Cumulus Cells ◽  
Treated Group ◽  
Vitro Fertilization ◽  
Oocyte Collection ◽  
First Results ◽  
Morula Stage ◽  
Serum Gonadotropin ◽  
Group 2 ◽  
Group 1

The Ban minipig is a local breed characterized by small ovaries with a scant number of follicles available for in vitro maturation (IVM). The combination of eCG and hCG has been used successfully to control estrus in pig breeding programs. In this paper we present the first results of IVF in this breed in comparison with 2 types of oocyte preparation: (1) from animals not receiving gonatotropin treatment (group 1, n = 9); and (2) from animals receiving an injection of mixed pregnant mare serum gonadotropin and hCG, 300 IU/animal, for 3 days before oocyte collection (group 2, n = 4). All animals were 1 to 3 years old and with body weights that varied from 8 to 12 kg. At the time of collection, the ovaries were observed for follicle development; the cumulus–oocyte complexes (COCs) were aspirated using a 18-gauge needle. COCs of categories A (with more than 4 layers of cumulus cells) and B (with 2 to 4 layers of cumulus cells) were collected and matured in vitro as described previously (Kikuchi et al. 2002 Biol. Reprod. 66, 1033–1041) at 39�C under 5% CO2 in air. Matured oocytes with expanding cumulus cells were inseminated using male Ban minipig epididymal semen frozen by the methods reported by Kikuchi et al. (1998 Theriogenology 50, 615–623). The frozen–thawed spermatozoa were pre-incubated for 1 h in modified medium-199 adjusted to pH 7.8 in the incubator at 37�C. The capacitated spermatozoa were diluted and added to drops of fertilization medium (Fig-FM; Suzuki et al. 2002 Int. J. Androl. 123, 135–142) containing oocytes; the final concentration of sperm was 106/mL. After 3 h of co-incubation, attached spermatozoa and cumulus cells were removed from oocytes and the oocytes were the cultured in vitro as described previously (Kikuchi et al. 2002). The results obtained from 4 replicates showed that the number of follicles with a diameter larger than 2 mm and the rates of oocytes categorized as A and B were significantly lower (P &lt; 0.05; ANOVA test) in the nontreated animals (0.0 and 67.5%, respectively) than in the treated group (25.5 and 87.1%, respectively). The rates of oocytes with a clearly expanding cumulus obtained after IVM were 78.6 (n = 136) and 88.1% (n = 101) for groups 1 and 2, respectively. The rates of cleaved embryos and embryos developed to the compact morula stage were 47.2 and 9.1% (n = 39), respectively, for group 1; and 89.1 and 18.8% (n = 101), respectively, for group 2. In conclusion, gonadotropin treatment before the collection of oocytes is recommended for application of IVM–IVF to local Ban minipigs. This work was supported by a grant from the VAST-Japan Society for the Promotion of Science Project.

307 ADDITION OF HIGH CONCENTRATIONS OF HEPARIN DURING IN VITRO FERTILIZATION IMPROVES PERCENTAGES OF CLEAVAGE AND BLASTOCYST FORMATION WHEN BOVINE OOCYTES ARE INSEMINATED IN VITRO WITH SEX-SORTED SPERM

2007 ◽  
Vol 19 (1) ◽  
pp. 269 ◽  
Author(s):  
T. L. Nedambale ◽  
J. Xu ◽  
S. A. Chaubal ◽  
X. C. Tian ◽  
X. Yang ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Optimal Concentration ◽  
Blastocyst Formation ◽  
Developmental Potential ◽  
Vitro Fertilization ◽  
High Concentrations ◽  
Bovine Oocytes

The developmental potential of oocytes fertilized in vitro by sexed–sorted frozen–thawed sperm remains low. The aim of this study was to find an optimal concentration of heparin during IVF using frozen–thawed sex-sorted sperm to improve the percentages of fertilization and blastocyst formation. A total of 1708 matured bovine oocytes were randomly allocated among different concentrations of heparin (0, 2.5, 5, 10, 20, and 40 µg mL−1) in Bracket-Olifant medium and co-incubated with non-sexed or sex-sorted sperm for 6 h (Day 0). The sperm (sorted or not) used for IVF was from one bull of proven fertility. Presumptive zygotes then were cultured in CR1aa + 6 mg mL−1 of BSA in 5% O2, 5% CO2, and 90% N2 at 39°C until Day 8. Percentages of cleavage were recorded on Day 2. Data (3 replicates) were analyzed by ANOVA. The results (Table 1) demonstrated that addition of different concentrations of heparin in IVF medium for non-sexed sperm groups did not alter cleavage and blastocyst formation. However, the lowest percentages of cleavage were observed with 0, 2.5, and 5 µg mL−1 of heparin in sexed sperm groups, demonstrating that high concentrations of heparin (20 and 40 µg mL−1) positively affected the percentages of cleavage and blastocyst formation. A greater proportion of zygotes in 20 and 40 µg mL−1 sexed sperm groups developed into grade 1 blastocysts on Day 7, and subsequently formed more blastocysts on Day 8 compared with 0, 2.5, and 5 µg mL−1 of heparin in sexed sperm groups. In conclusion, addition of heparin was not necessary when frozen–thawed non-sexed sperm were used for IVF with this particlar bull. However, addition of higher concentrations of heparin during IVF improved fertilization and blastocyst formation in vitro (40 µg mL−1 of heparin could be used as an alternative concentration for frozen–thawed sex-sorted sperm with this particular bull). The incidence of polyspermy as well as polyploidy in blastocysts is currently being studied. Table 1.Effect of heparin concentrations during IVF on cleavage and blastocyst formation

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