Conditional and cumulative live birth rates after blastocyst transfer.

OBJECTIVE: We aimed to investigate live birth rate (LBR), cumulative live birth rate (CLBR) for consecutive fresh and frozen-thawed in vitro fertilization (IVF) cycles, and CLBR after an entire IVF programme involving multiple ovarian stimulations using blastocyst transfer only. STUDY DESIGN: From January 1 st 2014 to December 31 st 2018, we included women aged 18-45 years who initiated IVF or intracytoplasmic sperm injection at Aagaard Fertility Clinic, Denmark. The primary outcome was live birth, and secondary outcomes were a positive hCG blood test and ongoing pregnancy confirmed by ultrasonography. All proportions were estimated for initiated and transferred cycles with 95 % confidence intervals (CI). We used a conservative strategy, assuming that none of the women who did not return for further treatments had a live birth. RESULTS: 871 women contributed 2236 initiated/1670 transferred fresh and/or frozen-thawed cycles. LBRs for first fresh cycles were 22.8 % (95 %-CI: 19.8-26.0) and 35.7 % (95 %-CI: 31.4-40.2) for initiated and transferred cycles, respectively. LBRs for first frozen-thawed cycles were 30.6 % (95 %-CI: 26.4-35.1) and 31.7 % (95 %-CI: 27.4-36.3) for initiated and transferred cycles, respectively. CLBRs for consecutive cycles were 18.2 % (95 %-CI: 16.2-20.3) for fresh initiated cycles, 29.7 % (95 %-CI: 26.6-32.9) for fresh transferred cycles, 25.5 % (95 %-CI: 22.6-28.5) for frozen-thawed initiated cycles, and 26.4 % (95 %-CI: 23.5-29.6) for frozen-thawed transferred cycles. For 436 women who contributed with an entire IVF programme we found a CLBR of 64.0 % (95 %-CI: 59.3-68.5). CONCLUSION: Compared to other studies of CLBR after multiple ovarian stimulations using cleavage stage transfer, our study presents a considerable effect in the IVF success rate when using blastocyst transfer only. In a clinical setting, transfer of blastocysts seems to be a viable method.

DNA Fragmentation in Human Spermatozoa and Pregnancy Rates after Intrauterine Insemination. Should the DFI Threshold Be Lowered?

Sperm DNA fragmentation index (DFI) can be analyzed by a flow cytometric assay after treatment with acid and acridine orange. In this prospective, cohort study, the value of DFI was determined in a semen analysis collected before fertility treatment (baselineDFI) in 146 couples and during 1-3 intrauterine inseminations (IUI) in 211 couples (511 cycles). The pregnancy rate (PR)/cycle was 9.9% if baselineDFI was >10 and 21.7% if baselineDFI was ≤10, (p < 0.005). The live birth rate (LBR)/cycle was 5% if baselineDFI was >10 and 14.2% if baselineDFI was ≤10 (p < 0.005). PR/patient was 23.1% if baselineDFI was >10 and 45.5% if baselineDFI was ≤10 (p < 0.005). LBR/patient was 12.4% if baselineDFI was >10 and 34% if baselineDFI was ≤10 (p < 0.005). When isolating non-stimulated IUI cycles and couples with female age < 35, a significant difference in PR and LBR between couples with high DFI and low DFI was seen. Results suggest that DFI > 10 could advice against timed coitus and non-stimulated IUI cycles. Analysis for DFI performed before treatment provides information about PR and LBR after IUI.

Implementation of an in-house flow cytometric analysis of DNA fragmentation in spermatozoa.

An increased amount of DNA fragmentation in the spermatozoa (SDF) is linked to male infertility. The Sperm Chromatin Structure Assay (SCSA) is widely used for analysis of SDF. However, the current software (SCSASoft®) linked to this assay is licensed and often located within larger diagnostic centers. In this study, we present a protocol for using other types of software than SCSASoft® to determine the SDF index (DFI) with clinical relevance. This protocol is engineered after collecting and analyzing 254 samples from fertility patients and sperm donors over a 15-month period. DFI is analyzed using a strict protocol where the spermatozoa are treated with a strong acid (pH 1.2) followed by acridine orange. DFI is determined by a standard flow cytometric software, FACSDiva 6.1.3. Analysis of the outcome of the fertility treatment is included for 137 patients receiving either intrauterine inseminations (IUI) or timed coitus (TC). The results show that the chance of pregnancy declines as DFI increases. We also found that the male DFI affects the chance of pregnancy independent of the female age. We have shown that a standard flow cytometric software can be used when determining a clinical relevant DFI. These findings are a significant step toward implementing the analysis as a part of the routine, in-house diagnosing of the male fertility patient and subsequently optimizing the treatment course of the couple with reduced human and financial costs.