Secretome profile selection of optimal IVF embryos by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

PURPOSE: Selecting an embryo at the transfer stage with the best chance of a successful pregnancy is still largely dependent on preceding subjective evaluation of morphokinetics. Expensive prenatal genomic profiling has been so far proved ineffective. Proteomics and metabolomics are promising new approaches to assess embryo viability, but methodologies are often complex and do not lend themselves to rapid analysis in the critical time between blastocyst formation and embryo transfer. Here, we used matrix-assisted laser desorption ionization time-of-flight (MALDI ToF) mass spectrometry to assess the secretome of blastocysts in the minutes prior to embryo transfer and correlated spectral features with pregnancy outcome. METHODS: Four hundred one samples of spent blastocyst culture media were collected from embryo cultures at the time of embryo transfer, of which 136 were used to construct the predictive model. The media samples were frozen at - 20 °C and stored for analysis. Sample analysis was conducted in batches using 1 µl of spent embryo in direct MALDI ToF mass spectral analysis. Quantitative characteristics within this mass range (2000-17,000 m/z) were used to generate a score for selected mass regions (bins) in order to predict pregnancy outcome for each sample. RESULTS: With a simple algorithm based on nine mass bins within the 2000-10,000 m/z region, it was possible to identify samples with the best chance of becoming an ongoing pregnancy (positive predictive value of 82.9%, p = 0.0018). CONCLUSION: A simple, direct and rapid analysis of spent culture fluid from blastocysts at the point of embryo transfer can quickly identify optimal embryos with the best chance of achieving ongoing pregnancy. Methods like this, which take less than 20 min to perform, could dramatically improve the approach to embryo selection and live births.

A prospective, randomized study comparing morning to evening administration of gonadotropins in ART.

OBJECTIVE: We sought to determine whether administering the daily gonadotropin dose in the morning (AM) or in the evening (PM) affects cycle outcome in patients undergoing IVF. DESIGN: This is a prospective randomized study. SETTING: The study is performed in a private assisted reproductive technology (ART) clinic. PATIENT(S): The study included one hundred and twenty-seven women undergoing IVF. INTERVENTION(S): Morning (AM) and evening (PM) administration of gonadotropins (uFSH and hMG) was compared. MAIN OUTCOME MEASURE(S): Live birth rate was the main outcome measured. Secondary outcomes including total IU use, days of stimulation, peak E2, peak P4, endometrial thickness, number of oocytes retrieved, MII oocytes, fertilization rates, #ET, IR, and clinical PR were all assessed. RESULTS: A total of 127 cycles were included, 61 in the AM group and 67 in the PM group. Baseline and stimulation characteristics were similar in both groups. There was a trend for a higher implantation rate in the AM group vs. the PM group (60.3 vs. 47.2%, P = 0.066). The AM group had a higher chemical pregnancy rate compared to the PM group (81.7 vs. 65.6%, P = 0.024) and a higher clinical pregnancy rate (78.3 vs. 62.1%, P = 0.048), but the delivery rates were similar (68.3 vs. 56.1%, P = 0.16). The study was unfortunately prematurely terminated when uFSH (Bravelle©) was pulled out of the US market. CONCLUSIONS: AM administration of gonadotropins may be associated with a better ART outcome compared to PM administration. Larger studies are needed to confirm our findings.

Decreased gonadotropin requirements in once daily compared to twice daily administration: a prospective, randomized study.

OBJECTIVE: Despite the lack of any scientific data, many ART programs split the daily gonadotropin dose during ovarian stimulation, while others give the entire dose during a single administration, usually at night. DESIGN: Prospective randomized. PATIENT(S): 213 women undergoing IVF/ICSI cycles at a single private ART center. INTERVENTION(S): Gonadotropin administration once daily compared to twice daily. MAIN OUTCOME MEASURE(S): Gonadotropin usage, clinical and ongoing PR RESULTS: There were 110 women in the once daily compared to 103 in the twice daily arm. All cycles were blastocyst transfers. There was a significantly lower FSH use in the once daily arm compared to the twice daily arm (1507.5 ± 517.5 IU vs. 1702.5 ± 622.5, P = 0.015), and a trend towards lower hMG use in the once daily arm (1342.5 ± 562.5 IU vs. 1462.5 ± 645.0, P = 0.15), without compromising clinical pregnancy rate (PR) (71.8% vs. 70.9%, P = NS) or delivery/ongoing PR (58.2% vs. 62.1%, P = NS). There were no differences in age, body mass index (BMI), peak estradiol, peak progesterone, retrieved oocytes, fertilized oocytes, number of ET, or PR. CONCLUSIONS: Once daily administration is associated with lower gonadotropin usage without compromising success rates.

Incidence of monozygotic twins in blastocyst and cleavage stage assisted reproductive technology cycles.

OBJECTIVE: To study the incidence of monozygotic twins (MZT) in blastocyst and cleavage stage ET. DESIGN: Retrospective review. PATIENT(S): Four hundred ninety-six women undergoing IVF/intracytoplasmic sperm injection cycles at a private assisted reproductive technology (ART) center. INTERVENTION(S): Patients undergoing ART were divided according to the stage of ET into blastocyst transfer (BT) and cleavage stage (CS) ET. MAIN OUTCOME MEASURE(S): Incidence of MZT as noted on vaginal ultrasound at 6 to 8 weeks. RESULT(S): There were 374 (75.4%) BT cycles, and 122 (24.6%) CS cycles. Women in the BT group were significantly younger, had more oocytes retrieved, and had less embryos transferred compared with the CS group. The clinical pregnancy rate was significantly higher in the BT group at 67.9% (254 of 374), compared with 37.7% (46 of 122) among the CS group. There was a significantly higher incidence of multiple gestation in the BT group compared with CS group (37.4% compared with 19.6%). The overall incidence of MZT was 1.3%, but differed with the stage of ET: the incidence of MZT was 1.57% (4 of 254) in the BT group, and 0 (0 of 46) in the CS group. CONCLUSION(S): Contrary to the older published literature on MZT in BT cycles, the incidence of MZT is low. Women undergoing ART therefore should not be discouraged to undergo BT for fear of MZT, especially in light of the higher pregnancy rate and lower number of transferred embryos noted in those cycles compared with cleavage stage transfers.