Non-invasive evaluation of embryos using mid-infrared attenuated total reflection spectrometry of incubation medium: a preliminary study.

RESEARCH QUESTION: Can mid-infrared attenuated total reflection (MIR ATR) spectroscopy combined with machine learning methods be used as an additional tool to predict embryo quality and IVF treatment outcomes? DESIGN: Spent culture media was collected and analysed. MIR ATR absorbance spectra were measured using an ALPHA II spectrometer equipped with an attenuated total reflection (ATR) spectrometry accessory. Patient and treatment data and results were collected and analysed in combination with machine learning techniques to identify possible correlations. The main outcome measures were to define the characteristics of absorbance spectra of spent culture media and to distinguish the difference in absorbance between top- and low-quality embryos, day 3 and day 5 embryos and implanting embryos versus non-implanting embryos. RESULTS: Spent culture media of 227 embryos was collected and analysed. Absorbance peaks in the culture media were different between day 3 and day 5 embryos. Moreover, significant differences in P-values, spanning from 0.014 to 0.044 in absorbance peaks for day 3 embryos and 0.024 up to 0.04 for day 5 embryos, were seen between implanting and non-implanting embryos. Machine learning techniques offered a pregnancy prediction value of 84.6% for day 3 embryos. CONCLUSIONS: MIR ATR may offer an additional parameter for better selection of embryos based on the spectrometric absorbance and secretions of metabolites in the culture media.

Can we alter pregnancy outcome by adjusting progesterone treatment at mid-luteal phase: a randomized controlled trial.

Our study aimed to determine whether mid-luteal serum P concentrations can serve as a predictive factor for in vitro fertilization (IVF) outcomes and whether increasing P dosage for patients with low levels at mid-luteal phase may improve pregnancy rates. It was a prospective, randomized controlled study. A total of 146 patients undergoing IVF treatment were prospectively enrolled and received routine luteal phase support (LPS) regimen of Endometrin® (progesterone) 200 mg/day. Serum P levels were measured 7 days after embryo transfer (ET). Considering a cutoff level of 15 ng/ml on this day, patients with higher levels continued the same dosage until pregnancy test (control group). Patients with lower levels were randomly allocated to continue Endometrin® 200 mg/day (Group A) or to increase Endometrin® dosage to 300 mg/day (Group B). The Main Outcome Measures were pregnancy rates. Both biochemical and clinical pregnancy and live birth rates were comparable between all groups regardless of P level on day 7 of luteal phase and regardless of dose adjustment. ROC analysis determined that mid-luteal P levels of 17 ng/ml can be a better predictor of cycle outcome. In conclusion raising the P dose at mid-luteal phase to 300 mg daily did not improve cycle outcomes.

Obesity results with smaller oocyte in in vitro fertilization/intracytoplasmic sperm injection cycles-a prospective study.

BACKGROUND: Obesity is associated with several fertility disorders. This prospective cohort study was designed to evaluate the effect of body mass index (BMI) (kg/m2) on oocyte diameter and treatment. METHODS: Women undergoing in vitro fertilization (IVF) with intracytoplasmic sperm injection (ICSI) were enrolled in the study. They were divided into two groups according to BMI: obese (BMI > 30) and normal weight (BMI < 25). Mature oocytes were evaluated according to total diameter, zona pellucida, and oolema diameters. RESULTS: A total of 387 oocytes were obtained from the 46 women who participated. Significantly more mature oocytes (M2) were retrieved from normal weight patients compare to obese women (15.1 ± 6.8 vs. 9.7 ± 3.9, respectively, P < 0.001). Oocytes from women in the obese group were significantly smaller than those in the normal weight group, including oocyte diameter (157.9 ± 7.9 vs. 164.3 ± 5.1 µm, P < 0.0001), oolema diameter (110.3 ± 4.5 vs. 113.5 ± 3.5 µm, P < 0.0001), and zona pellucida thickness (17.9 ± 2.6 vs. 19.0 ± 2.4 µm, P < 0.000), respectively. Multivariate logistic regression analysis, including oolema diameter, female age, BMI, number of M2 oocytes, and zona pellucida, was conducted to predict pregnancy. Small oolema diameter in obese patient adversely correlated with pregnancy. Larger oolema diameter was positively associated with the probability of pregnancy in the obese group as well as thinner zona pellucida. CONCLUSION: Obesity is associated with smaller oocytes, which adversely affect fertility outcomes. TRIAL REGISTRATION: NIH number NCT01672931.

Can intra cytoplasmatic morphologically selected sperm injection (IMSI) technique improve outcome in patients with repeated IVF-ICSI failure? a comparative study.

OBJECTIVE: Spermatozoal morphology was reported to effect fertilization, embryo quality and pregnancy results in spontaneous conception and ART. Intracytoplasmic morphologically selected sperm injection (IMSI) is an innovative, not invasive technique, which examines the sperm with no harm at a magnification of 6000 × in order to obtain optimal sperm to perform IVF-ICSI. We evaluated the efficiency of IMSI technique in patients with repeated IVF-ICSI failure of at least three cycles with no viable pregnancy and/or very poor sperm quality. STUDY DESIGN AND METHODS: All couples who performed IMSI between the years 2009 to 2012 were enrolled retrospectively to the study. Couples with male infertility who were treated with IMSI were included in the study. All their treatments were evaluated and divided into two subgroups: conventional IVF-ICSI treatment and their subsequent IMSI treatment. Demographic data, clinical parameters and outcome were recorded. The IMSI treatments were compared to previous non-IMSI treatments in terms of fertilization rates, cleavage rates, number of embryos and their quality, number of embryos transferred and pregnancy outcome. MAIN RESULTS: Forty-two couples were reviewed. Basic characteristics of the groups were comparable. Fertilization and cleavage rates of the two groups were comparable. The embryos quality demonstrated a trend toward superior quality (grade 1-2) embryos in the IMSI versus ICSI (60% versus 47%; p = 0.07 and 53% versus 40%; p = 0.07), respectively. Implantation and clinical pregnancy rates were significantly superior in IMSI group (19.2% versus 7.8%; p = 0.042 and 41.3% versus 10.5%; p = 0.02, respectively). Miscarriage rate was significantly higher in conventional IVF-ICSI group (100% versus 15.8%; p = 0.04), and live birth rate was significantly higher in IMSI group (0 in conventional IVF-ICSI and 34.7% per transfer in IMSI group; p = 0.003). CONCLUSION: IVF outcome of IMSI resulted in a higher implantation rate, pregnancy rate and most importantly delivery rate compare to non IMSI treated cycles.

[In vitro maturation of oocytes: a novel method of treatment of patients with polycystic ovarian syndrome undergoing in vitro fertilization].

BACKGROUND: In vitro maturation of oocytes (IVM) was developed to make in vitro fertilization (IVF) safer and simpler mainly for women with poLycystic ovarian syndrome (PCOS). The major benefits of IVM treatment include avoidance of hormone administration and risk of ovarian hyperstimulation syndrome (OHSS). OBJECTIVE: Results of IVM from our unit in patients with PCOS. METHODS: In this study 85 PCOS patients underwent 102 IVF cycles of IVM and were treated with one of the following protocols: (1) Priming with 150 units of recombinant FSH for 3 days, from the 3rd day of menses, following follicle development up to 10-12 mm. (2) Administration of 17beta estradiol (estrofem) on second day of menses, followed by ultrasound endometrial measurement up to > or =6 mm. thickness. Oocytes were collected 38 hours post recombinant human chorionic gonadotropin (rhCG) administration. Luteal phase support was achieved by estrofem and progesterone. Oocytes were matured either 6-30 hours (protocol 1) or 24-48 hours (protocol 2) in IVM medium and fertilized by intracytoplasmatic sperm injection (ICSI). Mean number of immature oocytes collected, maturation, fertilization, cleavage and pregnancy rates were assessed. RESULTS: Total number of retrieved oocytes was 1224 (mean 12 +/- 6.2 per cycle); 820 (64.9%) underwent maturation after 6-48 hours of culture while 128 of them (15.6%) after 6 hours and 20.2% of transferred embryos originated from those oocytes. FertiLization rate was 47.2%. Pregnancy and implantation rates were 28.4% and 11.25% respectively. CONCLUSIONS: IVM appears to be a simpler and tolerable treatment method in patients with PCOS undergoing IVF treatment. Favorable results were obtained.

CD11c+HLADR+ dendritic cells are present in human ovarian follicular fluid, and their maturity correlates with serum estradiol levels in response to gonadotropins.

OBJECTIVE: To determine whether dendritic cells (DCs), innate immune cells that specialize in initiation and modulation of immune responses, are present in ovarian follicular fluid (FF) and whether their abundance and maturation state correlate with ovarian response to gonadotropins. DESIGN: Observational study. SETTING: IVF unit and laboratory for reproductive immunology. PATIENT(S): Patients undergoing IVF. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): FF was collected from the first follicle aspirated in each patient, and cellular content was analyzed by flow cytometry. DCs were defined as CD45(+)CD11c(+)HLADR(+)-cells, and the intensity of HLADR expression indicated DC maturity. RESULT(S): The CD45(+)-hematopoietic cell compartment in FFs (n = 30) contained a significant fraction of CD11c(+)HLADR(+) DCs (15.4% ± 2.9%). The mean fluorescence intensity (MFI) of HLADR expression, which reflects DC maturity, correlated positively with ovarian response to gondotropins, as determined by serum levels of E(2) on the day of hCG administration (r = 0.38). CONCLUSION(S): DCs make up a significant fraction of hematopoietic cells in the FF. Furthermore, DC maturation correlates positively with the ovarian response to gonadotropins. It is therefore conceivable that DCs contribute to the sterile inflammatory process in the follicle that leads to ovulation.

Maturation of human ovarian follicles is accompanied by a decrease in the CD56+CD16+ natural killer cell population.

OBJECTIVE: To investigate whether "proangiogenic" CD56+CD16- natural killer (NK) cells, which accumulate in follicular fluid (FF) of patients with a good response to ovarian stimulation, are also present in earlier stages of follicular development. DESIGN: Observational study. SETTING: Academic in vitro fertilization (IVF) unit. PATIENT(S): Patients of similar age and ovarian reserve, undergoing in vitro maturation (IVM; n=10) or IVF (n=22) cycles. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): FF was collected from the first follicle aspirated in each ovary, and flow cytometry was used to define CD56+CD16- "proangiogenic" or CD56+CD16+ "cytotoxic" NK cells. RESULT(S): FF derived from antral follicles of patients undergoing IVM (maximum diameter 10 mm) showed a slightly higher abundance of "proangiogenic" NK cells compared with FF from preovulatory mature follicles (>18 mm) of patients undergoing IVF (5.4±1.3% vs. 3.0±1.1% of CD45+CD3- cells). Importantly, antral FF contained a significantly higher concentration of "cytotoxic" NK cells (11.4±2.3% vs. 3.7±0.9% of CD45+CD3- cells) compared with FF from mature follicles. CONCLUSION(S): "Proangiogenic" NK cells accumulate in ovarian follicles from as early as the antral follicular stage. Maturation of follicles is accompanied by a decrease in the population of "cytotoxic" NK cells that may have deleterious effects on follicular maturation.