Thyroid function and IVF outcome for different indications of subfertility

Studies evaluating pregnancy outcomes after assisted reproductive treatment (ART) in women with high-normal (2.5-4.5 mIU/L) TSH levels are conflicting, possibly due to different patient charactistics and subfertility indications. The aim of this study was to examine the hypothesis that high-normal compared to low-normal TSH levels are associated with adverse implications for pregnancy outcomes in conventional in vitro fertilization (IVF)-treated women. Therefore we analysed retrospectively the characteristics and pregnancy outcomes of 949 subfertile women with TSH 0.3-4.5 mIU/L, treated with conventional IVF between January 2008 and March 2012. Demographic and baseline characteristics were compared between groups of patients based on TSH quartiles, using one-way Anova, Kruskall-Wallis ANOVA and chi-square test. Women with high-normal quartile TSH were significantly more likely to be primary subfertile (p = 0.01), with a higher prevalence of unexplained subfertility and with 15% fewer live births after IVF compared to lower TSH quartiles (p = 0.02). In secondary subfertile women with high-normal TSH male factor subfertility prevailed (p=0.01), with more live births (p=0.01). When analysing primary and secondary subfertile women as one group, these differences failed to be observed, showing no differences in cumulative pregnancy outcomes of IVF between TSH quartiles (I: 0.3-1.21 mIU/L; II: 1.22-1.68 mIU/L; III: 1.69-2.31 mIU/L; IV: 2.32-4.5 mIU/L). In conclusion, primary subfertile women predominate in the high-normal TSH quartile, associated with significantly fewer live births in a subgroup of primary unexplained subfertile women (9%; n=87/949), while in secondary subfertile women, dominated by male factor subfertility, high-normal TSH is associated with more live births.

Download Full-text

252 COMPARISON OF TWO SPERM SEPARATION PRODUCTS FOR USE IN BOVINE IVF

Reproduction Fertility and Development ◽

10.1071/rdv17n2ab252 ◽

2005 ◽

Vol 17 (2) ◽

pp. 276 ◽

Cited By ~ 1

Author(s):

J. Pryor ◽

S. Romo ◽

D.D. Varner ◽

K. Hinrichs ◽

C.R. Looney

Keyword(s):

Sperm Concentration ◽

Embryo Production ◽

Chi Square ◽

Vitro Fertilization ◽

Before And After ◽

Chi Square Test ◽

Group 2 ◽

Live Sperm ◽

Sperm Separation

In commercial bovine in vitro fertilization (IVF) companies, there is a continuous need to improve results. Efforts to maximize in vitro embryo production have included modifications in the use of sperm separation gradients. The development of commercially available sperm centrifugation gradients represents a new possibility of increasing the number of viable sperm that can be obtained from low concentration (fresh or frozen, sexed or unsexed) semen samples in order to improve the efficiency of the IVF system to make embryo production as efficient as possible. The objective of this study was to compare two different separation gradients, as follows: Group 1: Percoll (Sigma, St. Louis, MO, USA), in 45% and 90% gradients; Group 2: EquiPure (Nidacon, Gathenburg, Sweden), in top and bottom layers. Before and after separation, sperm were evaluated at 200× magnification for total motility, and then stained to assess viability at 400× with fast-green/eosin stain (Sigma). Sperm separation was performed using frozen/thawed semen from one bull. Semen was separated by centrifugation at 200g for 30 min in both density gradients. Results obtained from Groups 1 and 2 were compared by chi-square test. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 92 × 106, vs. 159 × 106 sperm/mL for EquiPure; P < 0.05) but resulted in higher motility (60% vs. 39%, respectively; P < 0.05) of separated sperm. Rates of live sperm cells were not significantly different between groups (69.5% vs. 70%, respectively; P > 0.1). These results indicate that the commercial separation medium EquiPure may be associated with higher sperm concentration levels but with lowered sperm motility when compared to Percoll for bovine sperm separation. However, Equipure provided similar percentages of live sperm when compared to Percoll, which is currently used in our laboratory.

Download Full-text

Factors Related to the Number of Antral Follicles on InVitro Fertilization (IVF)

Indonesian Journal of Obstetrics and Gynecology ◽

10.32771/inajog.v4i2.79 ◽

2016 ◽

pp. 75

Author(s):

Fransiskus C Raharja ◽

Ketut Suwiyoga ◽

IPG Wardhiana

Keyword(s):

Prevalence Ratio ◽

Cross Sectional Study ◽

Chi Square ◽

Cross Sectional ◽

Vitro Fertilization ◽

Antral Follicles ◽

Chi Square Test ◽

The Mean ◽

Infertile Patients

Objective: To determine factors which are related to the number of antral follicles on infertile patients. Method: This cross sectional study was conducted in In-Vitro Fertilization (IVF) clinic of Graha Tunjung, Sanglah hospital, Bali. All fertile patients following the IVF program were calculated the number of antral follicles in both ovarian using transgene USG. This sample was recruited by random sampling from April 1st, 2001 to April 30th, 2011. We analyzed the data using Chi square test through SPSS for Windows 17.0 version. Result: Of 102 samples, the mean of patients’ age was 32.9% (SD 4.6) years old. From 72 patients (70.6%) experienced above 3 years of infertile period, the primary infertile was on 69 patients (67.7%). There was a relationship between patients’ age and the number of antral follicles significantly (prevalence ratio (PR) 1.41; 95% CI 1.11- 1.79). Meanwhile, the number of antral follicles and type of infertile (PR 1.02; 95% CI 0.76-1.37) also infertile period (PR 0.95; 95% CI 0.72-1.27) were not associated significantly. Conclusion: Patients’ age has an association with the number of antral follicles on IVF. [Indones J Obstet Gynecol 2016; 4-2: 75-77] Keywords: age, infertile, infertile period, number of antral follicles and type of infertile

Download Full-text

Predictive Factors For Three Pronuclear Zygote In Ivf Cycle

KnE Medicine ◽

10.18502/kme.v1i1.531 ◽

2016 ◽

Vol 1 (1) ◽

Author(s):

Budi Wiweko

Keyword(s):

Strong Predictor ◽

Multivariate Logistic Regression Analysis ◽

Controlled Ovarian Hyperstimulation ◽

Categorical Variables ◽

Continuous Variables ◽

Chi Square ◽

Preimplantation Genetic Testing ◽

Vitro Fertilization ◽

Chi Square Test

Introduction: Preimplantation Genetic Testing is one of the methods to screen genetic defects in embryos created through In Vitro Fertilization. In developing country this technique is still new and expensive method. Thus, knowing several factors in predicting the occurrence of three-pronuclear zygote can help much.Method: 472 cycles in 449 subjects who underwent controlled ovarian hyperstimulation in IVF cycles between January 2013 and August 2014 were included in the study. Categorical variables were compared using Chi Square test and continuous variables were analyzed using Independent t-test, and statistically significant was considered when p < 0.05. Multivariate logistic regression analysis was performed in order to correlate clinical variables and the occurrence of three-pronuclear zygote (3pn). Results: There were 38 3pn was identified in this study (8.05%). No correlation was found between age of the women, number of mature follicles, which are less than six with the incidence of 3pn. However, level of Anti Mullerian Hormones (AMH) found to be the strongest predictor with the incidence of 3PN (p < 0,01, RR 2.5, CI95% 1,55; 4.16).Conclusion: level of AMH is known to be strong predictor of thee pronuclear zygote after IVF cycle.

Download Full-text

188 EFFECT OF TRANSFER OF FROZEN-THAWED IVF EMBRYOS ON PREGNANCY IN REPEAT-BREEDER INSEMINATED OR NON-INSEMINATED HOLSTEIN CATTLE

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab188 ◽

2006 ◽

Vol 18 (2) ◽

pp. 202 ◽

Cited By ~ 2

Author(s):

O. Dochi ◽

M. Tanisawa ◽

S. Goda ◽

H. Koyama

Keyword(s):

Pregnancy Rate ◽

Embryo Transfer ◽

Calf Serum ◽

Pregnancy Rates ◽

Holstein Cattle ◽

Chi Square ◽

Vitro Fertilization ◽

Chi Square Test ◽

Repeat Breeder

Repeat-breeding is one of the important factors that affect dairy management. The objective of this study was to investigate the effect of transfer of frozen–thawed IVF embryos on pregnancy in repeat-breeder Holstein cattle. Cumulus–oocyte complexes (COCs) were collected by aspiration of 2–1-mm follicles from ovaries obtained at a local abattoir. COCs were matured for 20 h in TCM-199 supplemented with 5% calf serum (CS) and 0.02 mg/mL of FSH at 38.5°C under a 5% CO2 atmosphere in air. Matured oocytes were inseminated with spermatozoa of 5 × 106/mL in BO solution (Brackett and Oliphant 1975 Biol. Reprod. 12, 260–274) containing 10 mM hypotaurine and 4 units/mL heparin. After 18 h of gamete co-culture, presumptive zygotes were cultured in CR1aa (Rosenkrans et al. 1991 Theriogenology 35, 266) supplemented with 5% CS for 8 days at 38.5°C under 5% CO2, 5% O2, 90% N2 atmosphere in air. After in vitro fertilization, Day 7 and Day 8 blastocysts were frozen in 1.5 M ethylene glycol (EG) in Dulbecco's PBS (DPBS) supplemented with 0.1 M sucrose and 20% CS. Embryos were transferred into a freezing medium, loaded into 0.25-mL straws, and allowed to stand for 15–20 min for equilibration. The straws were then plunged into a −7°C methanol bath of a programmable freezer for 1 min, seeded at −7°C, maintained at −7°C for 15 min, cooled to −30°C at the rate of −0.3°C/min, and then plunged into liquid nitrogen. Recipient animals (43 heifers, 131 cows) included those that did not conceive after being artificially inseminated (AI) 3 to 15 times. The frozen–thawed IVF embryos were directly transferred to the recipient animals 7 days after estrus or AI. Pregnancy rates were analyzed by chi-square test. The results are presented in Table 1. There were no significant differences in the pregnancy rates between treatments. However, a slightly higher pregnancy rate was achieved by embryo transfer after AI. These results suggest that embryo transfer may increase the pregnancy rate in repeat-breeder Holstein cattle. Table 1. Pregnancy rates after transfer of IVF frozen–thawed embryos in repeat-breeder Holstein cattle

Download Full-text

Evaluation of comparison of clinical outcome treated oocyte with ionomycin and strontium chloride in infertile couple with previous failed in vitro fertilization

Journal of Shahid Sadoughi University of Medical Sciences ◽

10.18502/ssu.v27i2.1047 ◽

2019 ◽

Author(s):

Marziyeh Norozi-Hafshejani ◽

Marziyeh Tavalaee ◽

Mohammad Hossein Nasr- Esfahani

Keyword(s):

Clinical Results ◽

Fertilization Rate ◽

Infertile Couple ◽

Strontium Chloride ◽

Oocyte Activation ◽

Male Factor ◽

Chi Square ◽

Vitro Fertilization ◽

Failed Fertilization

Introduction: Several factors are involved in failed fertilization following intracytoplasmic sperm injection (ICSI), which could be related to sperm, oocyte factors and/or both. Failure in oocyte activation is considered as the most important factor in failed fertilization after ICSI. To overcome this shortcoming, artificial oocyte activation (AOA) after ICSI has been suggested. Commonly, ionomycin and strontium chloride are used as the most efficient agents for oocyte activation in the clinic. In this study, for the first time, the clinical results of ionomycin and strontium chloride were compared on sister oocytes of couples who previously have had fertilization failure after ICSI. Methods: In this intervention study, 14 couples with male factor infertility and previous failed fertilization after ICSI were included in this study. Oocytes of their wives were divided into two groups. Half of the oocytes after ICSI were treated with ionomicin and other half were treated with strontium chloride. Then, fertilization rate, embryo quality and pregnancy status were compared through SPSS 18 software, Paired sample t-test and chi-square test. Results: Percentage of fertilization rate was significantly higher in oocytes that were activated using strontium chloride compared to those that were activated with ionomycin (p<0.0001). Although, the pregnancy rate was insignificantly higher in strontium chloride group than ionomycin group (p=0.387), but the quality of embryo was similar between the two groups (p=0.924). Conclusion: In infertile men with previous failed fertilization after ICSI, the use of strontium chloride can also be recommended for oocyte activation.

Download Full-text

124 CO-CULTURE WITH BOVINE INTACT CUMULUS - OOCYTE COMPLEXES DURING IN VITRO FERTILIZATION OF BUFFALO DENUDED OOCYTES COMPLETELY RESTORES THEIR FERTILIZING AND DEVELOPMENTAL COMPETENCE

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab124 ◽

2011 ◽

Vol 23 (1) ◽

pp. 167

Author(s):

M. De Blasi ◽

M. Rubessa ◽

L. Boccia ◽

S. Di Francesco ◽

M. V. Suárez Novoa ◽

...

Keyword(s):

Cumulus Cells ◽

Blastocyst Stage ◽

Negative Control ◽

Developmental Competence ◽

Control Group ◽

Chi Square ◽

Oocyte Vitrification ◽

Vitro Fertilization ◽

Chi Square Test

Removal of cumulus cells is necessary for several technologies such as vitrification, intracytoplasmic sperm injection, and nuclear transfer. However, it is known that the presence of cumulus cells during IVF of buffalo oocytes is fundamental for fertilization and embryo development (Gasparrini et al. 2007 Anim. Reprod. Sci. 98, 335–342; Nandi et al. 1998 Theriogenology 50, 1251–1262). The aim of this work was to evaluate whether co-culture with intact bovine cumulus–oocyte complexes (COC) during IVF would restore the developmental competence of denuded buffalo oocytes. Due to the scarce availability of buffalo ovaries, the somatic support was provided by bovine cumulus cells. Abattoir-derived COC were matured in vitro according to our standard procedures (Gasparrini et al. 2006, Theriogenology, 65, 275–287) and randomly distributed in 3 fertilization groups: 1) a control group of COC (n = 122), 2) a negative control of denuded oocytes (DO; n = 119), and 3) DO co-cultured with in vitro matured bovine COC (DO+COC; n = 103) in a 1:1 ratio (3 bovine COC + 3 denuded buffalo oocytes/50 μL drop). Fertilization was carried out with frozen–thawed spermatozoa from a tested bull in TALP medium supplemented by 0.2 mM penicillamine, 0.1 mM hypotaurine, and 0.01 mM heparin at 38.5°C under a controlled gas atmosphere of 5% CO2 in humidified air. After fertilization the zygotes were cultured in SOF medium including essential and nonessential amino acids and 8 mg mL–1 BSA, at 38.5°C under humidified 5% CO2, 7% O2, and 88% N2, up to the blastocyst stage. On Day 5 and on Day 7 (Day 0 = IVF) cleavage and blastocyst rates were respectively recorded. Data were analysed by chi-square test. As expected, cleavage and blastocyst rates were lower (P < 0.01) in DO (36.1 and 9.2%, respectively) compared with the control (67.2 and 27.1%, respectively). However, co-culture during IVF (DO+COC) significantly increased (P < 0.01) both parameters compared with DO, giving cleavage (70.9%) and blastocyst (27.2%) rates similar to the control. The results of this study demonstrated that co-culture with bovine intact COC during IVF of buffalo denuded oocytes completely restores their fertilizing capability and blastocyst developmental competence. We conclude that this may be a suitable strategy for preserving the developmental competence of oocytes devolved to technologies, such as oocyte vitrification, that require cumulus removal.

Download Full-text

202 EFFECT OF OSTEOPONTIN ON IN VITRO EMBRYO DEVELOPMENT IN CATTLE

Reproduction Fertility and Development ◽

10.1071/rdv20n1ab202 ◽

2008 ◽

Vol 20 (1) ◽

pp. 180 ◽

Cited By ~ 1

Author(s):

B. Gasparrini ◽

E. Monaco ◽

L. Boccia ◽

A. De Rosa ◽

L. Attanasio ◽

...

Keyword(s):

Embryo Development ◽

Bovine Milk ◽

Cumulus Cells ◽

Control Group ◽

Single Chain ◽

Chi Square ◽

Vitro Fertilization ◽

Chi Square Test ◽

Oviduct Fluid

Osteopontin (OPN) is an acidic single-chain phosphorylated glycoprotein found both in the oviduct fluid (ODF) and oviductal epithelium in cattle, which is believed to facilitate fertilization. It was recently reported that addition of a rabbit polyclonal immunoglobulin G antibody against purified bovine milk OPN with sperm oocytes, bovine oocytes, or both decreased (P < 0.05) fertilization compared with the in vitro-fertilized control (Goncalves et al. 2007 Theriogenology 67, 468–74). The aim of the present work was to determine the effect of in vitro addition of OPN to the fertilization medium on both cleavage and postfertilization embryo development in cattle. In the first experiment, in vitro-matured oocytes were fertilized in modified TALP medium in the presence of 0.0, 0.1, 1.0, or 10 µg mL–1 of OPN. In a second experiment, matured oocytes were in vitro-fertilized in modified TALP medium in the presence of 0.0, 10, 20, or 40 µg mL–1 of OPN. In vitro fertilization was carried out with frozen–thawed spermatozoa from a bull previously tested for IVF. After 20 to 22 h of coincubation at 39�C, 5% in CO2 in air, presumptive zygotes were vortexed to remove cumulus cells, washed, and transferred, 30 to 50 per well, into 400 µL of SOF modified medium. Zygotes were incubated in a humidified mixture of 5% CO2, 7% O2, and 88% N2 in air at a temperature of 39�C. On Day 7 of development (Day 0 = day of insemination), cleavage and development rates into transferable embryos (TE)–tight morulae (TM) and blastocysts (Bl) of superior quality were recorded. Differences in the percentages of both cleavage and blastocyst rates among groups were analyzed by a chi-square test. In experiment 1, numerically higher percentages of TM–Bl (29.5, 29.5, 30.5, and 37.5%, respectively, in the control group and in the groups with 0.1, 1, and 10 µg mL–1 of OPN; P = 0.25) and Bl (28.6, 27.5, 29.1, and 36.7, respectively, in the control group and in the groups with 0.1, 1, and 10 µg mL–1 of OPN; P = 0.24) were observed with 10 µg mL–1 of OPN. In experiment 2, significantly more cleavage (80.0 v. 71.3%; P < 0.05) and higher percentages of TM–Bl (44.6 v. 34.5%; P < 0.05) and Bl (39.2 v. 30.6%; P = 0.06) were observed with 10 µg mL–1 of OPN v. the control. Combined analysis from both experiments showed an overall effect of 10 µg mL–1 of OPN v. the control in the percentages of TM–Bl and Bl (respectively, 41.1 v. 33.3% and 37.7 v. 30.5%; P < 0.05). These results indicate that it is possible to improve the efficiency of bovine in vitro embryo production by adding the oviductal protein OPN.

Download Full-text