Preimplantation genetic diagnosis: does embryo biopsy for PGD compromise clinical outcome?

ence of GSK-3␤ in non-gonadal embryonic mouse tissues and genital ridges of day 12.5 p.c female sex embryos. The adult rat testis, known to test positive for GSK-3␤ by IHC, served as a positive control. A negative control was performed by omitting primary antibody to GSK-3␤. Embryos were harvested at day 12.5 after a noticeable vaginal pug is detected in a mated adult female mouse. Embryos were sexed through a previously published PCR technique detecting expression of Sry in male sex embryos. Ongoing studies detail the expression of GSK-3␤ in earlier and later dates of female embryogenesis and meiotic progression. Results: Strong expression was demonstrated in the brain in the mesencephalon and in the dorsal root ganglia and spinal cord mantle. In the liver there is moderate interspersed expression in canalicular cells and hepatocytes. In addition there was moderate staining in cardiac myocytes and endothelial cells. Finally, moderate staining could also be shown in mesonephric and metanephric tubules and interspersed cells of the gonadal ridge. Conclusions: These findings are the first to report a detailed account of GSK-3␤ in female non-gonadal and gonadal tissues and support a role for this kinase in the development of the central and peripheral nervous systems, the liver heart and kidneys. Its expression in the gonadal ridge at time of onset of meiosis strengthens the evidence for a role of this kinase in the early events of meiotic initiation. P-36 Preimplantation genetic diagnosis: does embryo biopsy for PGD compromise clinical outcome? N. Ouhibi1, P.E. Patton,1 K.A. Burry,1 S. Olson,2 H. Lawce,2 M.J. Gorrill,1 D.M. Lee,1 D.E. Battaglia.1 Depts. of 1 OB/GYN and 2Molecular and Medical Genetics, Oregon Health & Science University, Portland, OR. Background: Preimplantation genetic diagnosis (PGD) is a technique to screen for chromosomal and genetic disorders in embryos before pregnancy has been established. This technology is now recognized as a tool for the diagnosis of inherited and chromosomal disorders in the clinical IVF setting. Objective: The aims of this study were (1) to compare the clinical outcome between PGD and IVF patients and (2) to evaluate the developmental competence of normal and chromosomally abnormal embryos. Design: This was a prospective study of IVF and PGD patients undergoing blastocyst transfer after embryo biopsy. Pregnancy, implantation and blastocyst rates were compared between the PGD group (n ⫽ 38) and the age-matched control IVF group (n ⫽ 36). All patients that had embryos available received an embryo transfer on d5 or d6. Materials and Methods: Embryo biopsy was performed in Ca2⫹/Mg2⫹ -free medium on all embryos with 5 cells or greater on day-3. Biopsied blastomeres were subjected to fluorescent in situ hybridization (FISH) for chromosomes 13, 18, 21, X and Y (PGT probes from Vysis). Embryos were then cultured to the blastocyst stage. Significant differences (p ⬍ 0.05) were analyzed using Chi square test or Fisher Exact test. Results:

Conclusions: Fertilization rate, blastocyst rate, implantation rate, and pregnancy rate/embryo transfer were not significantly different between the two groups. However, pregnancy rates/cycle were significantly lower in PGD couples secondary to the high degree of aneuploidy in biopsied embryos resulting in no transfer. These data support the principle that the biopsy procedure does not compromise pregnancy potential.

P-37 Midluteal estradiol-to-progesterone ratio (E2/P4) has no effect on IVF outcome. I. Souter,1 D. Hill,2 M.W. Surrey.3 1Dept. of Ob/Gyn, David Geffen School of Medicine at UCLA, Los Angeles, CA, 2ART Center and 3 Southern California Reproductive Center, Beverly Hills, CA. Background: Progesterone secretion is considered the major hormonal event during the luteal phase, and estradiol appears to play a crucial role as well. Whether an optimal estradiol-to-progesterone ratio is critical for implantation and progression of early pregnancy remains unclear. Objective: To determine whether altered midluteal E2/P4 ratios have an effect on embryo implantation and pregnancy maintenance in IVF cycles. Materials and Methods: We retrospectively evaluated 97 patients undergoing IVF-fresh ET. Serum was obtained 5 days after ET and concentrations of estradiol and progesterone were determined. The ratio of estradiol-toprogesterone was calculated for conception and non-conception cycles. Data are expressed as mean ⫾ SD. Student’s t-test, ␹2 and Fisher’s exact test were used for statistical analysis. Results: 97 IVF-ET cycles resulted in 53 pregnancies (54.7%). Fifty-eight day-3, sixteen day-4, and twenty-three day-5 ETs resulted in 29,7, and 17 pregnancies respectively. Clinical pregnancy rates were 41.3%, 43.8%, and 60.9% respectively. Five days after ET ongoing clinical pregnancies had significantly higher progesterone values compared to spontaneous abortion and non-conception cycles (80.3 ⫾ 56 ng/dL, 64.2 ⫾ 36 ng/dL, and 59.3 ⫾ 31 ng/dL respectively- p:0.0425). This effect was more pronounced five days after day-3 ETs (92.2 ⫾ 62 ng/dL vs 59.6 ⫾ 33.5 ng/dL-p:0.025). Progesterone concentrations higher than 60 ng/dL were associated with significantly increased pregnancy rates (62.5% vs 34.6%-p:0.035). Five days after a day-3 ET, higher E2/P4 ratios were noted in unsuccessful cycles compared to clinical pregnancy ones (12.6 vs 8.2, p:0.08). A trend towards a negative effect on pregnancy rates was noted in all cycles (day-3 to day-5 ETs), when the ratio was ⬎15.0 (PR:30%, p: NS), however these observations did not reach statistical significance. Evaluation of different E2/P4 ratios did not reveal any significant differences in pregnancy and implantation rates between conception and non-conception cycles (Fig.1).

Table 1: Clinical outcome: Age-matched comparison: PGD GROUP

IVF GROUP

P VALUE

NO. OF CYCLES 38 36 NS MEAN AGE 38 37.2 NS TRANSFER RATE (%) 22/38* 35/36** 0.0001 FERTILIZATION 359/522 (68.7) 401/546 (73.4) NS RATE (%) BLASTOCYST 105/292 (35.9) 130/381 (34.10) NS RATE (%) NO. OF EMBRYOS 40 86 NS TRANSFERRED NO. OF SACS 8 22 NS IMPLANTATION RATE (%) 20 25.6 NS PREGNANCY RATE/ 21 36 0.0291 CYCLE (%) PREGNANCY RATE/ 36.4 37 NS EMBRYO TRANSFER (%) * 16 patients did not have a transfer, of these 3 had no blastocyst development and 13 had chromosomally abnormal embryos. ** 1 patient did not get a transfer (no blastocyst).

FERTILITY & STERILITY威

Conclusions: Our data suggest that altered midluteal E2/P4 ratios do not result in reduced implantation and pregnancy rates. However, midluteal progesterone concentration of ⬎60ng/dl is associated with increased endometrial receptivity and successful IVF outcome. Luteal phase support should be directed towards optimizing progesterone concentration without a goal of altering the balance of estradiol to progesterone, since this is not crucial for implantation and pregnancy maintenance.

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