Live births after IVF in men with a DNA fragmentation index of 30% or greater as determined by the sperm chromatin structure assay (SCSA™)

Monday, October 18, 2004 5:15 P.M. O-116 Live births after IVF in men with a DNA fragmentation index of 30% or greater as determined by the Sperm Chromatin Structure Assay (SCSA™). F. Barnes, F. Rabara, A. Murphy, C. Zouves. Zouves Fertility Center, Daly City, CA. OBJECTIVE: Evaluate the implantation and birth frequency in couples where the male partner had a DFI of ⱖ 30 %. DESIGN: Retrospective review of IVF cycles with a DNA fragmentation index (DFI) ⱖ 30%. MATERIALS AND METHODS: Semen samples provided by men undergoing IVF were subjected to SCSA™ testing within 0 –180 days of IVF. Samples were produced by masturbation into specimen cups (Mckesson Medi-pak #37–2469P) and processed within 15–300 minutes. Samples were evaluated for count and motility and frozen neat and stored for one to fourteen days in liquid nitrogen Dewars. Frozen samples were sent to SCSA™ Diagnostics, Inc, Brookings, SD for testing. Results were available within 7 days. Men with a high DFI (ⱖ 30%) were advised to use Fertility Blend™ supplement for men and couples were counseled to add an additional embryo to the transfer if available. In vitro fertilization (IVF), embryo culture and embryo transfer were performed by processes standard to the industry. Sperm donation was not suggested as an option for IVF treatment. Data presented includes men with DFI’s ⱖ 30%, pregnancy and birth data where derived from only the first cycle following SCSA testing. Data is grouped in thirty-day intervals from SCSA testing to egg recovery. RESULTS: Men with high DFI’s were consistently able to produce healthy children following in vitro fertilization regardless of the time of testing to egg retrieval.

OBJECTIVE: The Sperm Chromatin Structure Assay (SCSA), a newly available diagnostic tool for IVF practitioners, measures the percentage of sperm with high levels of DNA fragmentation ⫺ DNA Fragmentation Index (% DFI). The results are used to ascertain the fertility potential of patients receiving fertility treatment. The objective of this study was to evaluate the correlation between DFI and outcome in patients receiving ICSI treatment. DESIGN: A retrospective analysis of 68 IVF patient cycles, between January 2003 and April 2004 with SCSA test results. MATERIALS AND METHODS: Patients were included if they received ICSI, a day 3 transfer and did not have pre-implantation genetic diagnosis. Patients were divided into 3 groups depending on DFI score: * Excellent fertility potential ⬃ ⫺/⬍15% DFI (Group I) * Good fertility potential ⫺/⬍ 15–30% DFI (Group II) * Poor fertility potential ⬃ ⬎30% DFI (Group III) The embryos were scored on day 3 accounting for morphology, cell number, and cell fragmentation, using a grading system whereby 4 is highest quality and 1 is poorest quality. A statistical analysis using one-way ANOVA and CHI-SQUARED test was used to determine any significant trends in rates of fertilization, embryo quality and pregnancy (⫹hCG) with a P level of 0.05. RESULTS: Of the total population, 19 patients were in group I, 29 in group II and 20 in group III. Fertilization rates in groups I, II and III were 62.2 (⫾18.7), 60.9 (⫾20.8) and 50.7 (⫾20.7) respectively (P⬎0.05). Average embryo quality on day 3 in groups I, II and III was 2.83 (⫾0.82), 2.97 (⫾0.59) and 2.98 (⫾0.58) respectively (P⬎0.05). Pregnancy rates in groups I, II and III were 10/19 (53%), 16/29 (55%) and 9/20 (45%) respectively (P⬎0.05). The difference in female age between the 3 groups was not significant; the average age in groups I, II and III was 34.2 (⫾5.46), 33.9 (⫾4.75) and 33.95 (⫾4.40) respectively (P⬎0.05). The number of embryos transferred between the 3 groups was not significant; the average number transferred in groups I, II and III was 2.74 (⫾0.93), 2.48 (⫾0.87) and 3.25 (⫾1.773) respectively (P⬎0.05). CONCLUSION: This analysis of initial data shows no significant difference between all groups in fertilization rate (although there does seem to be a trend towards decreasing fertilization rate with increasing DFI), embryo quality on day 3 or pregnancy rate. An extension of the present study may be warrented to strengthen the value of these findings by increasing the size of the study groups, and improving standards of protocol, for example performing the SCSA analysis on the day of oocytes retrieval, removing time factors and effects of any treatment (such as anti-oxidant therapy). Most importantly, these results do show that even with a high DFI score, patients can still achieve a pregnancy, including one patient with a DFI of 49 who has an ongoing twin pregnancy. Supported by: None

Monday, October 18, 2004 5:45 P.M. O-118

1: Clinical pregnancy defined as the presence of a fetal sac at 6 weeks. 2: Sacs electively terminated prior to birth. Added note: three patients with DFI’s ⱖ50%, within 4 and 59 days of egg retrieval produced 7 healthy children following IVF. CONCLUSION: The data presented is in contrast to reports that a DFI ⱖ 30% strongly predicts a male patient’s inability to produce a sustained pregnancy. Fertility Blend™ supplement for men contains L-Carnitine and other antioxidants that may have led to reduced and improved DFI scores by the time of IVF. Men with an initial DFI ⱖ 30% should not move immediately to sperm donation when attempting to father a child by IVF. Supported by: None

Monday, October 18, 2004 5:30 P.M. O-117 A high DFI score does not preclude patients from achieving a pregnancy with ICSI. T. A. Elliott, T. H. Taylor, C. A. Jacobs, W. E. Roudebush, W. Brockman, Z. P. Nagy. Reproductive Biology Associates, Atlanta, GA.

FERTILITY & STERILITY威

Clinical outcome of intracytoplasmic sperm injection (ICSI) with frozen-thawed epididymal and testicular sperm with an investigation of chromosomal abnormalities in the sperm and ICSI pregnancies. S. Ma, H. Gao, S. S. Tang, B. Ho Yuen, V. Chow, M. Nigro. University of British Columbia, Vancouver, BC, Canada. OBJECTIVE: The aims of the study are: 1) to measure the efficacy of ICSI, in terms of clinical outcome, with frozen-thawed epididymal and testicular spermatozoa, 2) to correlate chromosomal abnormalities in the frozen-thawed spermatozoa with the clinical outcome, and 3) to investigate cytogenetically the newborn babies conceived from this procedure. DESIGN: Prospective study MATERIALS AND METHODS: In 31 patients (17 suffering from obstructive azoospermia, OA, 9 from non-obstructive azoospermia, NOA, 5 from failed microsurgical reversal for vasectomy), sperm were retrieved from either microsurgical epididymal sperm aspiration (MESA) (n⫽20) or testicular biopsies (TESE) (n⫽11). Cryopreserved spermatozoa from the day of operation were frozen-thawed and used for 41 planned ICSI cycles. Spermatozoa leftover from 10 ICSI procedures (5 from MESA and 5 from TESE) were processed for FISH, after patients’ consent. Probes for chromosomes X, Y and 18 were used to detect chromosomal abnormalities in the patient sperm, as well as in sperm from 5 proven fertile men (controls). Chromosomal analysis of cord blood from newborn babies was also carried out. RESULTS: Fertilization rates were similar in MESA and TESE groups

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