Larry I. Barmat

Expression of Inhibin/Activin Subunits and Their Receptors and Binding Proteins in Human Preimplantation Embryos

Purpose:Our purpose was to study the role of inhibin/activin during embryogenesis.Methods:Transcripts of inhibin/activin subunits (α, βA, βB), activin receptors (types I and II), and follistatin were detected by a reverse transcriptase–polymerase chain reaction in human reproductive cells and preembryos cultured alone or co-cultured with human endometrial cells.Results:Transcripts of α, βA, βBsubunits were all detected in granulosa luteal cells, but only βAunits were detected in endometrial stromal and decidualized cells. In human preimplantation embryos, none of these subunits were detected in embryos from the four-cell to the morula stage and only βAsubunits were detectable in blastocyst embryos. Activin receptors were detectable in all of the studied embryos and cells. Transcripts of βA, activin receptors, and follistatin were differentially expressed in human preimplantation embryos cultured in vitro and their expressions were significantly enhanced with the presence of endometrial stromal cells.Conclusions:Our data suggest that there is a possible endometrium–embryo interaction via endometrial activins and preimplantation embryo receptors and that the embryonic expressions of these activins, their receptors, and binding proteins are dependent on embryonic stage.

The embryo toxicity of hydrosalpinx fluid is only apparent at high concentrations: an in vitro model that simulates in vivo events

OBJECTIVE To simulate the in vivo model in studying the effect of hydrosalpinx fluid on embryonic development. DESIGN Controlled prospective study. SETTING Academic research center. PATIENT(S) Five hundred eighty-seven two-cell murine embryos. INTERVENTION(S) Embryos were grown under two sets of conditions. Half were cultured using 10% fetal calf serum in RPM1 medium in varying concentrations of hydrosalpinx fluid (0, 1%, 10%, 50%, 75%, and 100%). To more closely mimic the in vivo environment, the other half were grown in an endometrial coculture system with the same media and hydrosalpinx fluid concentrations. MAIN OUTCOME MEASURE(S) Embryonic development. RESULT(S) For each stage of embryogenesis, diminished development was noted with increasing concentrations of hydrosalpinx fluid. In the group of embryos grown without endometrial coculture, only at a minimum concentration of 50% hydrosalpinx fluid was diminished development noted for the blastocyst, hatching, and outgrowth stages. When an endometrial coculture system was used, development was not inhibited until exposure to a minimum of 75% hydrosalpinx fluid. Embryogenesis was enhanced when an endometrial coculture system was used for each concentration of hydrosalpinx fluid. CONCLUSION(S) When a model is used that more accurately mimics the in vivo conditions of IVF-ET in a patient with hydrosalpinges, it appears that high concentrations of hydrosalpinx fluid are required to signiticantly impede embryogenesis. The endometrium appears to help detoxify hydrosalpinx fluid.

Human preembryo development on autologous endometrial coculture versus conventional medium

OBJECTIVE To evaluate the effect of autologous endometrial coculture versus conventional medium on preembryo development. DESIGN Controlled systematic clinical study. SETTING University-based IVF center. PATIENT(S) Women with a history of failed IVF-ET with poor preembryo quality. INTERVENTION(S) Patients underwent a luteal phase endometrial biopsy. The tissue then was digested enzymatically, and the stromal and glandular cells were separated by differential sedimentation rates. These cells were cultured to confluence, released, and then cryopreserved until the patients IVF-ET cycle. All normally fertilized oocytes then were allocated systematically to growth on autologous endometrial coculture or conventional medium until transfer on day 3. MAIN OUTCOME MEASURE(S) Preembryo blastomere numbers and cytoplasmic fragmentation rates were measured. RESULT(S) Forty-two women underwent 44 cycles of IVF-ET. In the morning on day 3, the mean (+/-SD) number of blastomeres and cytoplasmic fragments per preembryo on coculture compared with conventional medium was 5.9+/-1.5 versus 5.5+/-1.4 and 21%+/-13% versus 24%+/-11. At transfer the mean (+/-SD) number of blastomeres per preembryo on coculture was 7.4+/-1.3 versus 6.7+/-1.9 on conventional medium. CONCLUSION(S) There was a significant improvement in the mean (+/-SD) number of blastomeres per preembryo and decrease in the fragmentation rate for preembryos on autologous endometrial coculture compared with noncocultured preembryos from the same patient.

Autologous Endometrial Co-culture in Patients with Repeated Failures of Implantation After In Vitro Fertilization–Embryo Transfer

Purpose:Our purpose was to evaluate the effect of co-culture on preembryo development and clinical outcome.Methods:Enrolled patients underwent a luteal-phase endometrial biopsy. The tissue was then enzymatically digested (collagenase) and the stromal and glandular cells were separated by differential sedimentation rates. These cells were cultured to confluence, released, and then cryopreserved until the patients in vitro fertilization (IVF)–embryo transfer (ET) cycle. All normally fertilized oocytes were then placed on the co-cultured cells until transfer on day 3. Preembryo development on co-culture was compared to that in the patients noncocultured previous cycle. Implantation and clinical pregnancy rates were compared to those in a control group of patients undergoing IVF during the study period who were matched for age, stimulation protocol, number of oocytes retrieved, and preembryos transferred.Results:Twenty-nine women underwent 31 cycles of IVF-ET. On day 3 the overall mean number of blastomeres per preembryo on co-culture compared to that in the patients previous cycle was 6.3 ± 1.8 vs. 5.6 ± 1.2 (P = 0.04). The average percentage of cytoplasmic fragments on co-culture compared to the previous cycle was 16 ± 9% vs. 19 ± 9% (P = 0.32). At transfer, after preembryo selection, the mean number of blastomeres per preembryo on co-culture compared to that in the patients previous cycle was 6.8 ± 1.6 vs. 6.6 ± 1.3 (P = 0.5). The implantation and clinical pregnancy rates between co-culture and the matched control group were 15% (14/93) vs. 13% (16/124) (P = 0.79) and 29% (9/31) vs. 25% (10/40) (P = 0.45).Conclusions:There was a significant improvement in the average number of blastomeres per preembryo on co-culture compared to that in the patients previous noncoculture cycle. The overall implantation and clinical pregnancy rates between co-culture and a matched control group were not significantly different.

The effect of hydrosalpinges on IVF-ET outcome.

Purpose:Our purpose was to determine if the presence of a hydrosalpinx effects the outcome of in vitro fertilization (IVF)–embryo transfer.Methods:We performed a retrospective analysis of IVF cycle stimulation sheets.Results:A total of 1000 patients with tubal factor infertility was analyzed. There were 60 hydrosalpinx patients who underwent 116 initiated cycles with 106 embryo transfers, compared to 940 control patients undergoing 1428 initiated cycles with 1150 embryo transfers. Both groups had a similar response to ovarian stimulation, number of oocytes retrieved, and number of embryos transferred. The hydrosalpinx group had a significantly higher preclinical loss rate (22/59 = 37% vs 80/566 = 14%; P = 0.001), a significantly lower implantation rate (55/352 = 16% vs 795/3795 = 21%; P = 0.013), a trend toward a reduced delivery rate per transfer (28/106 = 26% vs 387/1150 = 34%; P = 0.066), a significantly higher ectopic pregnancy rate (5/59 = 8% vs 16/566 = 3%; P = 0.04), and a similar spontaneous abortion rate (9/37 = 24% vs 99/486 = 20%; P = 0.28) compared to the control tubal factor group.Conclusions:This study demonstrates a decrease in implantation rates and an increase in preclinical miscarriages and ectopic pregnancies in patients with hydrosalpinges compared to tubal-factor patients without sonographic evidence of dilated fallopian tubes.

Large, comparative, randomized double-blind trial confirming noninferiority of pregnancy rates for corifollitropin alfa compared with recombinant follicle-stimulating hormone in a gonadotropin-releasing hormone antagonist controlled ovarian stimulation protocol in older patients undergoing in vitro fertilization

OBJECTIVE To compare corifollitropin alfa with recombinant FSH treatment in terms of the vital pregnancy rate in older patients undergoing IVF. DESIGN Phase 3 randomized, double-blind, noninferiority trial. SETTING Multicenter trial. PATIENT(S) A total of 1,390 women aged 35-42 years. INTERVENTION(S) A single injection of 150 μg of corifollitropin alfa or daily 300 IU of recombinant FSH for the first 7 days then daily recombinant FSH until three follicles reach ≥17 mm in size. Ganirelix was started on stimulation day 5 up to and including the day of recombinant hCG administration. If available, two good quality embryos were transferred on day 3. MAIN OUTCOME MEASURE(S) Vital pregnancy rate (PR), number of oocytes, and live birth rate. RESULT(S) Vital PRs per started cycle were 23.9% in the corifollitropin alfa group and 26.9% in the recombinant FSH group, with an estimated difference (95% confidence interval) of -3.0% (-7.4 to 1.4). The mean (SD) number of recovered oocytes per started cycle was 10.7 (7.2) and 10.3 (6.8) in the corifollitropin alfa and the recombinant FSH groups, respectively, with an estimated difference of 0.5 (-0.2 to 1.2). The live birth rates per started cycle were 21.3% in the corifollitropin alfa group and 23.4% in the recombinant FSH group, with an estimated difference (95% confidence interval) -2.3% (-6.5 to 1.9). The incidence of serious adverse events was 0.4% versus 2.7% in the corifollitropin alfa and recombinant FSH groups, respectively, and of ovarian hyperstimulation syndrome (OHSS; all grades) was 1.7% in both groups. CONCLUSION(S) Treatment with corifollitropin alfa was proven noninferior to daily recombinant FSH with respect to vital PRs, number of oocytes retrieved, and live birth rates, and was generally well tolerated. CLINICAL TRIAL REGISTRATION NUMBER NCT01144416.

Importance of the biospy date in autologous endometrial cocultures for patients with multiple implantation failures

OBJECTIVE To analyze the effectiveness of autologous endometrial coculture by the cycle day of the endometrial biopsy. DESIGN Retrospective study. SETTING University-based IVF center. PATIENT(S) Two hundred eight patients with multiple IVF failures. INTERVENTION(S) Embryos were split and randomly allocated to growth on autologous endometrial coculture or conventional media. MAIN OUTCOME MEASURE(S) Embryo quality and pregnancy outcome. RESULT(S) The overall clinical pregnancy rate was 41.8%. Embryos grown on autologous endometrial coculture were of higher quality (more blastomeres and less fragmentation) than embryos grown with conventional media. Early luteal biopsies (<5 days after LH surge) for autologous endometrial coculture did not demonstrate an improvement in embryo quality as compared to the significant improvement demonstrated with later luteal endometrial biopsies (> or =5 days after LH surge). The date of the biopsy was predictive of pregnancy outcome when using autologous endometrial coculture (44.7% [> or =5 days after LH surge] vs. 18.8% [<5 days after LH surge], P=.012). CONCLUSION(S) We have demonstrated an improvement in embryo quality when using autologous endometrial coculture. The improvement in embryo quality and higher pregnancy rates were limited to biopsies > or =5 days after the LH surge. This suggests that mid/late luteal phase endometrium contains factors that enhanced embryo growth and subsequent implantation.

Oocyte donation: does a previous attempt affect a subsequent attempt?

OBJECTIVE To analyze the effect of a previous donor oocyte cycle on the outcome of subsequent attempts. DESIGN Retrospective study. SETTING Oocyte donation program at The New York Hospital/Cornell Medical Center. PATIENT(S) Two hundred sixty-seven patients undergoing 354 fresh cycles of oocyte donation. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Clinical outcomes were divided into groups based on the attempt number of each cycle for each patient. Results were calculated for each recipient cycle. RESULT(S) A clinical pregnancy rate of 56.2% and ongoing pregnancy/delivery rate per retrieval of 50.3% were noted. No statistically significant differences in clinical outcomes were found between the first, second, and third attempts. A significant increase was noted in the ongoing pregnancy/delivery rate per recipient cycle for the second attempt in those patients who had a delivery after the first attempt compared with those who did not. CONCLUSION(S) We demonstrated an overall clinical pregnancy rate of 56.2% and an ongoing pregnancy/delivery rate of 50.3% per retrieval. Outcome for the second attempt was associated with success or failure during an initial attempt at oocyte donation.

Autologous endometrial coculture in patients with a previous history of poor quality embryos.

AbstractPurpose: To evaluate the effect of autologous endometrial coculture in patients (less than 36 years old) with a history of a single IVF failed cycle associated with poor quality embryos. Methods:Design: Controlled clinical study. Setting: University-based in vitro fertilization center. Patients: Twenty-six patients with a history of a single prior failed IVF-ET with poor preembryo quality. Intervention(s): Autologous endometrial coculture. Main outcome measures: Preembryo blastomere numbers and cytoplasmic fragmentation rates were compared between the treatment and previous cycle. Clinical pregnancy rates were analyzed. Results: Twenty-six women with an average age of 32.8 ± 2.9 years underwent treatment. On Day 3 the overall mean number of blastomeres per preembryo on coculture compared to conventional medium in a previous cycle was 6.1 ± 1.8 vs. 5.1 ± 1.3 (P = 0.01; Wilcoxon test). The average percentage of cytoplasmic fragments on coculture compared to the conventional medium in a previous cycle was 14% ± 10 vs. 22% ± 13 (P = 0.003; Wilcoxon test). At transfer the mean number of blastomeres per preembryo on coculture was 7.4 ± 1.8 compared to 6.7 ± 1.5 on conventional medium in a previous cycle (P = 0.02; Wilcoxon test). The clinical pregnancy rate (positive fetal cardiac activity) per patient was 88.5%. The delivery rate was 73.1% (19/26). Conclusions: There was an improvement in the preembryo quality for preembryos on autologous endometrial coculture compared to noncocultured preembryos from the same patient in a previous cycle. An excellent delivery rate was subsequently found.