Faculty Bibliography

Cancer patients produce similar numbers of oocytes after ovarian hyperstimulation compared with age-matched infertile controls, suggesting that malignancy does not adversely affect ovarian response

In an 8-year review of ectopic pregnancy (EP) rates in donor egg recipients and standard patients undergoing in vitro fertilization-embryo transfer (IVF-ET) at a large university-based program, we report an EP rate of 0.6% in donor egg recipients and 0.9% in standard IVF patients, a difference that is not statistically significant. Donor egg recipients were found to have a significantly lower incidence of tubal disease compared with standard IVF patients; however, tubal disease was not found to be an independent risk factor for EP in our practice, perhaps owing to aggressive management of tubal disease

OBJECTIVE: Since many frequent, fatal, and incurable diseases do not have an appropriate disease model and with attainable embryos being scarce, with IRB approval we attempted to generate novel disease-specific human embryonic stem (hES) cells with new protocols for more efficient derivation, maintenance, and differentiation. DESIGN: Experimental. MATERIALS AND METHODS: Zona-free human blastocysts (n=14) previously assessed by preimplantation genetic diagnosis (PGD) for genetic conditions were transferred onto feeder cells and cultured in DMEM-based media. Pieces of the colonies (n=4) were frozen by liquid nitrogen vitrification with cryoprotectants propylene glycol, DMSO, and acetamide and subsequently thawed. Differentiation of hES cells was achieved by colony overgrowth or embryoid body formation. Embryonic and control cells were subjected to marker gene and protein analysis for pluripotency and differentiation by reverse transcription PCR and immunofluorescence, respectively. RESULTS: Colonies (n=10; 71.4% of transferred blastocysts) could be established and maintained which showed the typical morphological features of hES cells such as compact colony formation. Colonies were derived from affected embryos (one each of cystic fibrosis, trisomy X, 18, 21, 22, and Tay-Sachs disease), from embryos tested inconclusively (one of cystic fibrosis and three of Tay-Sachs disease), and from three normal control embryos. Marker gene and protein expression as well as growth pattern analysis suggest that the colony cells retain their undifferentiated state in culture as well as after vitrification and thawing and that they can be differentiated into a variety of cell types, including the tissues most affected by the conditions. CONCLUSIONS: These newly established protocols for the derivation, maintenance, and differentiation of novel disease-specific hES cell lines should enable the efficient generation of new disease models. This will provide new tools to study diseases as well as to develop new therapeutic approaches

OBJECTIVE: The use of OC is increasing. We know the human oocyte meiotic spindle (MS) is crucial to chromosome segregation and embryo development while the zona pellucida (ZP) aids in sperm binding and fertilization. Today, polarized microscopy (PM) affords non-invasive MS and ZP evaluation. All MII oocytes cryopreserved in our lab undergo PM MS and ZP assessment 2 h post-harvest. We compared OC cycle outcome in cases performed for MED vs. DR vs. OD. DESIGN: Retrospective. MATERIALS AND METHODS: All women were <40 y at time of OC. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS & 1 sarcoma. All patients underwent standard ovarian stimulation followed by retrieval, PM for MS and ZP, then OC of MII oocytes. RESULTS: Cycle outcome is shown in the Table. (Table presented). CONCLUSIONS: The majority of retrieved oocytes in all groups were MII. Women diagnosed with malignancy achieved adequate ovarian stimulation, egg production and a similar number of spindle positive eggs compared to DR and OD patients, despite limited time for treatment and multiple outside stressors, including underlying disease. MS & ZP measurements were clinically comparable whether eggs were frozen for cancer, DR or OD

OBJECTIVE: The MS is critical to chromosome segregation and embryo competence. Today, polarized light microscopy (PM) affords non-invasive MS evaluation (MSE). We perform PM 2 h post-harvest and 1 h post-thaw. Here, we appraise its value. DESIGN: Retrospective. MATERIALS AND METHODS: 28 OC cycles with thaw were assessed; 1/2 eggs were slow frozen and 1/2 vitrified (vit); pt's eggs were split between methods. MSE was done in the last 23 cycles. Outcome measures included 2PN fertilization (2PN-F), embryo quality suitable for ET, blast formation rate (BFR), and embryo usability (transferred or refrozen). RESULTS: All eggs were from women <38 (mean 31) yowith normal D3 FSH/E2. Of 28 cycles, 57% had pregnancy; 12 delivered 17 liveborns, 3 are ongoing and 1 aborted. In the last 25 cycles, 286 MII eggs were thawed after cryo for a mean of 3 mos; pre-cryo, 83% were MS positive (SP+). 255 (89%) survived; 89% were SP+ post-thaw, a 94% pre-cryo and post-thaw MSE correlation. 211 (83%) ICSI'd eggs achieved 2PN-F. 2.3 embryos were transferred/cycle (30 slow & 22 vit). Pre-cryo MSE: no difference was noted in egg survival or 2PN-F. 65% of SP+ vs. 44% of MS negative (SP-) zygotes became embryos suitable for ET D3 post-thaw (p=.02). No difference was noted in the BFR or % of embryos suitable for D5 ET, but 86% of embryos transferred or refrozen were from SP+ eggs. Post-thaw MSE: no difference was noted in the % of embryos suitable for ET D3 post-thaw or BFR. A significantly higher % of embryos were suitable for ET D5 post-thaw in the SP+ (57%) vs. SP- (29%) groups (p=.03) and 95% of transferred or refrozen embryos were from SP+ oocytes. When comparing MSE data for slow vs. vit eggs, no differences were noted. CONCLUSIONS: Most retrieved oocytes (slow and vit) exhibited a MS pre & post-thaw and most transferred or refrozen embryos were from SP+ oocytes. OC does not appear to damage the MS and viewing it may correlate with oocyte competence. Because MSE may improve efficiency of gamete selection, it should be considered to advance the field of OC

OBJECTIVE: As more reproductive-age women are diagnosed with and survive cancer, OC use is increasing as a means of FP, particularly in those without a male partner. We compared outcomes of medically-indicated (MED) OC cycles to those of infertile women completing an OC thaw cycle (THAW), including their meiotic spindle evaluation (MSE). All MII oocytes cryopreserved in our lab undergo MSE 2 h post-harvest. DESIGN: Retrospective. MATERIALS AND METHODS: 31 MED (520 retrieved eggs) and 28 THAW (512 retrieved eggs) OC cycles were reviewed. Tumors in the MED group included 13 GYN, 10 breast, 3 leukemia, 2 colon, 2 CNS and 1 sarcoma. All patients underwent standard ovarian hyperstimulation followed by retrieval, MSE 2 h post-harvest, then OC of all MII oocytes. Both slow freezing and vitrification were used. 4 MED pts froze ¹?₂ as zygotes. RESULTS: In the THAW cycles, there were 16 (57%) pregnancies: 12 women delivered 17 liveborns (5 twins), 3 have ongoing gestations and 1 miscarried. 90% of eggs survived and there was a 94% pre-cryo to post-THAW MSE correlation. 2 MED thaw cycles resulted in 1 pregnancy. Cycle comparisons for THAW vs. MED are shown in the Table. (Table presented). CONCLUSIONS: The majority of oocytes in both groups were mature (MII) and exhibited a spindle. Women diagnosed with cancer can achieve adequate ovarian stimulation, egg production and MSE despite limited time to complete OC treatment and multiple outside stressors, including their underlying disease and its management. Based on our preliminary THAW data, we believe cancer pts should have a reasonable chance for pregnancy using their frozen eggs

This study assessed 1908 embryos, including those with abnormal numbers of pronuclei, in IVF cycles from July 2001 to December 2006 in which preimplantation genetic screening (PGS) was performed on day 3 post-retrieval and 'euploid' embryos transferred the following day. PGS-intracytoplasmic sperm injection and PGS-translocation cycles were excluded. At 18 h post-insemination, the zygote distribution was 19% 0PN, 4% 1PN, 69% 2PN and 8% 3PN. No pregnancy occurred following 0PN or 1PN embryo transfers. A retrospective, blinded morphological ranking of all embryos on day 3 was performed and the results compared with PGS; no 0PN or 1PN embryo would have been chosen for transfer based on morphological superiority alone. Blastocyst formation occurred in 1PN embryos (29%) but not in 0PN embryos when evaluated on day 5. Euploid karyotypes were reported for biopsies of 0PN (3%), 1PN (5%) and 2PN (19%) embryos (P = 0.015, 1PN versus 2PN). A Y chromosome was observed in 0PN (17%) and 1PN (32%) embryos; surprisingly, 91% of these Y chromosome-bearing embryos were aneuploid. Many different meiotic and fertilization errors can result in 0PN or 1PN zygotes; these results indicate that the resultant embryos should not be transferred, especially when normally fertilized embryos are available

The intent of this study was to evaluate a recent randomized clinical trial evaluating the effect of preimplantation genetic screening (PGS) that reports a negative effect on pregnancy outcome. This article reviews appropriate PGS techniques and how they differ from the trial in question. A closer look at the clinical trial in question reveals significant lack of expertise in biopsy, cell fixation, genetic analysis, and patient selection. At most, this trial demonstrates that in inexperienced hands, PGS can be detrimental. No other conclusions concerning the effect of PGS on pregnancy results can be drawn from the trial.

We describe our experience of over 300 cycles of preimplantation genetic diagnosis (PGD) and report clinical pregnancy rates (35%-67%) that support using this technology to screen for genetic disorders and chromosomal abnormalities. In clinical practice for over ten years, PGD offers couples the earliest form of genetic screening and may help improve ongoing pregnancy rates in poor-prognosis patients