Rescue ICSI and PGD: the Combination Can Result in Pregnancy

Materials and Methods: This is a case report of a woman affected with SCA3. The patient’s mother had developed SCA3, so DNA-based testing was performed on the patient revealing 73 CAG repeats on one of the alleles of the MJD gene confirming the diagnosis of SCA3. In an effort to avoid passing the rare gene to her offsprings, PGD analysis was performed specifically selecting for SCA3 on the MJD gene. Results: The patient underwent two IVF cycles. In the first cycle, 11 oocytes were aspirated, and ICSI (Intracytoplasmic Sperm Injection) was performed on the 6 that appeared mature. On PGD analysis, 2 embryos yielded inconclusive results, 3 embryos were affected with SCA3, and 1 embryo was not affected however displayed monosomy 22. Therefore, no normal embryos were identified on the first cycle and no embryo transfer took place. In the second cycle, 18 oocytes were aspirated, 10 of which appeared mature for ICSI. Of these, 5 were affected, 3 were unable to be studied and 2 were normal after PGD analysis. These 2 embryos were subsequently transferred on day 6. Pregnancy was confirmed 10 days after the transfer and currently the patient carries a normal twin pregnancy. Conclusions: A specific gene was selected for with PGD analysis in IVF and embryos not carrying the selected mutation were selected resulting in normal twin pregnancy. PGD can be utilized to select for specific gene mutations known to be present in the family. P-53 Rescue ICSI and PGD: the Combination Can Result in Pregnancy. M. Li, MD, PhD,1, C. Marin De Ugarte, MD2, B. Jordan, BS1, M. Surrey, MD1, A. DeCherney, MD2, D. Hill, PhD1. 1ART Center, Beverly Hills, CA. 2UCLA Medical Center, LA, CA. Background: ICSI (Intracytoplasmic Sperm Injection) is usually performed in couples with male factor infertility on the day of the oocyte retrieval. PGD (Preimplantation Genetic Diagnosis) is performed for single gene disorders or aneuploidy testing. Objective: To describe a case in which embryos were treated by rescue ICSI in combination with PGD from which a pregnancy resulted. Materials and Methods: A case report is described. The patient is a 43 year old Gravida 2 Para 0020 with infertility with her present partner of one and half year duration. Her past OB/GYN history is significant for a spontaneous abortion with a biochemical pregnancy in 1988, as well as a therapeutic abortion in 1987. Her husband is a healthy 35 years old with previous proven fertility with a different partner several years ago. His semen analysis was obtained 8 months prior to the current IVF cycle and was deemed normal. The patient was stimulated in the following manner. Oral contraceptives were administered, followed by ovarian stimulation with gonadotropins along with a GnRH Antagonist. HCG was administered when follicle maturity was determined and 12 oocytes retrieved 36 hours later. On the day of retrieval, specimen collection from the patient’s husband resulted in 2.4ml of semen with 70 million/ml and 50% sperm motility rate. Oocytes were inseminated 4 h after retrieval. Results: On the following day no fertilization was observed and rescue ICSI was therefore performed on the 7 mature eggs, of which 2 normally fertilized. The patient desired PGD and therefore single blastomeres were then biopsied from 2 embryos 4 days after retrieval. FISH analysis was performed for chromosomes 13, 18, 21, X and Y. One embryo was diagnosed as normal and the other with trisomy 21. On day 5 the normal embryo was a 10 cell stage and was transferred into the patient’s uterus. The patient was continued on Progesterone supplementation and 2 weeks later the patient’s pregnancy test was positive with an hCG level of 560 mIu/ml. An intrauterine singleton pregnancy was then confirmed several weeks later by sonography. Conclusion: Rescue ICSI in combination with PGD can result in successful pregnancy. P-54 Day 3 Biopsy of Slow Cleaving, Poor Grade Embryos for Aneuploidy Screening Does Not Improve the Chance of Pregnancy in IVF-ET Patients. D. Hill1, M. Li1, C. Marin DeUgarte2, M. Surrey1, H. Danzer1, A. DeCherney2. 1ART Center, Beverly Hills, CA. 2UCLA Medical Center, LA, CA. Background: PGD (preimplantation genetic diagnosis) for the purpose of aneuploidy screening (AS) has increased in usage over the past decade and

FERTILITY & STERILITY威

is usually performed 3 days after oocyte retrieval. Our ART program currently performs AS on approximately twenty percent of patients requiring IVF-ET, and embryo biopsy was typically performed on virtually every embryo generated from these patients, regardless of their rate of cleavage or “grade” at biopsy. Objective: The aim of the study was to determine if slow cleaving, poor grade embryos are more likely to be chromosomally abnormal, and if biopsy of these embryos is either of value or is in fact a waste of medical resources. Materials and Methods: A retrospective analysis was performed on data collected from consecutive patients requesting aneuploidy screening of their embryos over a five month period. Morphological grading of embryos as well as their PGD results was recorded. Results: Out of 683 embryos biopsied from one hundred and eleven patients, 113 (16%) were ⬍6 cells at the time of biopsy on day three. Seventy four of 113 (or 65%) of these embryos were aneuploid. Further analysis of these embryos revealed a roughly equal distribution of normal versus aneuploid (26 vs. 30) outcomes if the grade of the embryos was in the A-B (good) range. In contrast, slow-cleaving embryos in the C-D (poor) morphologic range were five times as likely (11 normal vs. 55 aneuploid) to be abnormal in their chromosomal makeup, as well as three times as likely to have anucleated blastomeres (16 of 22) as did good grade, albeit slowcleaving embryos. Conclusion: This data suggests it is reasonable to biopsy ⬍6 cell embryos on day three post-insemination if they are of good morphology. Slowcleaving, morphologically compromised embryos are almost always aneuploid, and as these embryos invariably arrest upon extended culture, not performing AS on this subset of embryos poses little risk of discarding healthy, euploid embryos capable of establishing a sustained pregnancy, and has the added benefit of conserving valuable medical resources.

P-55 The Evaluation of Two Different Types of Sequential Culture Media Systems for Human In Vitro Fertilization Using Sister Oocytes. J. Chesmore, H. Lambert, A. Lopez, K. Vu. Hawaii Center for Reproductive Medicine and Surgery, Honolulu, Hawaii. Background: The development of commercial sequential culture media designed for human embryo culture has led to improvements in IVF and ICSI outcomes. These media are formulated according to the carbohydrate composition of oviduct and uterine fluids and take into account the changing physiology and metabolic requirements of the human embryo. Objective: To evaluate two sequential culture media systems in ART (Medicult® and Vitrolife®) using sister oocytes to measure fertilization rates (IVF and ICSI), cleavage and blastocyst development. Materials and Methods: Twenty three randomized patients with at least one failed IVF attempt were entered into the study under a standardized controlled ovulation stimulation (COS) protocol (Ortho-Cept 28®, Gonal F® Multidose, Cetrotide® 0.25mg., Ovidrel® and Crinone®). During this blinded study, sister oocytes were randomly selected after EPU and put into the two different types of sequential culture systems being evaluated. Embryos were examined daily during the 5 days of culture. Results: A total of 23 patients participated in the study with an average age of 34.1⫾3.2 years and an average FSH value of 6.8⫾1.1. The number of oocytes retrieved per patient was 18.7⫾9.5. Sequential Culture Medium Tested

Medicult®

Vitrolife®

P value

Avg.# Oocytes per Media Fertilization Rate (%) Day 3 Cleavage Rate (%) Compaction Rate (%) Morula Formation Rate (%) Blastocyst Formation Rate (%)

8.8⫾4.8 76.1⫾ 93.5⫾ 26.9⫾ 33.7⫾ 40.7⫾

8.9⫾4.8 72.8⫾ 82.5⫾ 17.6⫾ 23.2⫾ 19.7⫾

0.7458 0.0249 0.1751 0.1168 0.0020

Conclusion: Initially, there were no significant differences during fertilization however from day 3 to blastocyst, Medicult® significantly increased cell division and promoted blastocyst development and formation. Although numbers are small, the compelling data further suggests that HLA supple-

S31