Ryan M. Riggs

Does storage time influence postthaw survival and pregnancy outcome? An analysis of 11,768 cryopreserved human embryos

OBJECTIVE To evaluate the impact of cryopreservation storage duration on embryo survival, implantation competence, and pregnancy outcome. DESIGN Retrospective study. SETTING Academic tertiary-referral infertility center. PATIENT(S) In vitro fertilization patients and recipients of oocyte donation cycles who had cryopreserved embryos and underwent at least one thaw cycle from 1986 to 2007. INTERVENTION(S) None. MAIN OUTCOME MEASURE(S) Postthaw survival proportion and implantation, clinical pregnancy, miscarriage, and live birth rates. RESULT(S) Length of storage time did not have a significant effect on postthaw survival for IVF or oocyte donation cycles, or for embryos frozen at the pronuclear or cleavage stages. There was no significant impact of the duration of storage on clinical pregnancy, miscarriage, implantation, or live birth rate, whether from IVF or oocyte donation cycles. Logistic regression analysis demonstrated that the length of storage time or developmental stage at freezing were not predictive of embryo survival or pregnancy outcome. Only oocyte age, survival proportion, and number of transferred embryos were positive predictors of pregnancy outcome. CONCLUSION(S) Cryostorage duration did not adversely affect postthaw survival or pregnancy outcome in IVF or oocyte donation patients.

Assessment of ovarian reserve with anti-Müllerian hormone: a comparison of the predictive value of anti-Müllerian hormone, follicle-stimulating hormone, inhibin B, and age.

OBJECTIVE The objective of this study was to evaluate basal anti-Müllerian hormone as a marker for ovarian responsiveness to fertility treatment. STUDY DESIGN Frozen basal menstrual cycle day 3 serum samples were evaluated retrospectively for anti-Müllerian hormone, inhibin B, and follicle-stimulating hormone levels in 123 in vitro fertilization cycles (93 patients) and compared with in vitro fertilization records. RESULTS Anti-Müllerian hormone values correlated the best with the number of retrieved oocytes (r = 0.539; P < .001) relative to age (r = -0.323; P < .01), follicle-stimulating hormone (r = -0.317; P < .01), inhibin B (P > .05), luteinizing hormone (P > .05), and estradiol (r = -0.190; P < .05). Receiver operating characteristic curve analysis demonstrated that, for the prediction of <4 oocytes retrieved, anti-Müllerian hormone had the largest area under the curve (AUC = 0.81; P = .0001) relative to age (r = 0.74; P = .005), follicle-stimulating hormone (0.71; P = .02), inhibin B (0.66; P = .03), and estradiol (0.54; P > .05). Similarly, for the prediction of >or=15 retrieved oocytes, anti-Müllerian hormone had the largest area under the curve (0.80; P = .0001) relative to age (0.63; P = .02), follicle-stimulating hormone (0.64; P = .005), inhibin B (r = 0.57; P > .05), and estradiol (0.58; P > .05). CONCLUSION Anti-Müllerian hormone correlates better than age, follicle-stimulating hormone, luteinizing hormone, inhibin B, and estradiol with the number of retrieved oocytes. Receiver operating characteristic curves estimated that anti-Müllerian hormone accurately predicts ovarian responsiveness to controlled ovarian stimulation with high sensitivity and specificity.

Anti-Müllerian hormone serum levels predict response to controlled ovarian hyperstimulation but not embryo quality or pregnancy outcome in oocyte donation

The objective of this retrospective cross-sectional study was to evaluate the value of basal serum anti-Müllerian hormone (AMH) levels as a predictor of ovarian response and pregnancy outcome in a donor egg program. The study showed that AMH was superior to other biomarkers of ovarian reserve in predicting low and high response in young women selected as oocyte donors, but that it was not predictive of embryo morphology or pregnancy outcome in the recipient population.

Expression of milk fat globule EGF-factor 8 (MFG-E8) mRNA and protein in the human endometrium and its regulation by prolactin

Milk fat globule epidermal growth-factor 8 (MFG-E8) has not been previously linked to endometrial physiology. We reported on MFG-E8 mRNA up-regulation in the human endometrium during the window of implantation (WOI) using microarrays. Prolactin (PRL) secreted by stromal cells has been suggested to modulate protein expression. The objective of this study was to characterize the endometrial expression of MFG-E8 and its ligand αvβ3 integrin during the menstrual cycle and its possible regulation by PRL. MFG-E8 mRNA (real-time RT-PCR) and protein expression (immunohistochemistry and immunoblotting) were analyzed in human endometrial biopsies at different times of the menstrual cycle, as well as in primary endometrial cell cultures. In primary cultures of epithelial cells, MFG-E8 intracellular protein expression was evaluated in absence or presence of PRL (0.2 and 1 μg/ml). The results show that MFG-E8 protein is almost exclusively localized to the epithelium in whole endometrial biopsies. Both MFG-E8 mRNA and protein expression increased in the luteal phase and were highest during the WOI; epithelial protein location of αvβ3 integrin also peaked on cycle Day 24. Cultured epithelial cells showed a diffuse staining of MFG-E8 over the cytoplasmic area; however, some cells presented a punctuated staining pattern. PRL treatment of epithelial cells for 72 h in vitro significantly increased MFG-E8 protein intracellular expression. This is the first report on MFG-E8 protein localization to the human endometrial epithelium and its up-regulation during the WOI. The pattern of glandular expression of its ligand αvβ3 integrin was remarkably similar. In vitro data support a modulatory role for PRL as a stromal/epithelial paracrine factor controlling MFG-E8.

Epithelial cell protein milk fat globule-epidermal growth factor 8 and human chorionic gonadotropin regulate stromal cell apoptosis in the human endometrium.

OBJECTIVE To study the regulation of apoptosis in human endometrial cells. The specific aims were to determine whether milk fat globule-epidermal growth factor 8 (MFG-E8), a novel endometrial epithelial protein, modulates caspase activation and DNA fragmentation; and to examine whether hCG, an early embryonic product, regulates Bax and Bcl-2 equilibrium, as well as MFG-E8 expression. DESIGN Primary cultures of human endometrial epithelial cells (EECs) and endometrial stromal cells (ESCs). SETTING Academic center. PATIENT(S) Ovulatory women aged 21-30 years. INTERVENTION(S) Treatment with MFG-E8 and hCG. MAIN OUTCOME MEASURE(S) Apoptotic activity was quantified using a luciferase assay. Deoxyribonucleic acid fragmentation was detected by TUNEL assay. Bax, Bcl-2, and MFG-E8 messenger RNA expression levels were determined by quantitative reverse transcription-polymerase chain reaction. Immunocytochemistry was used to establish cell purity and presence of MFG-E8 and hCG-R (receptor) proteins. RESULT(S) Endometrial epithelial cells were cytokeratin(+), vimentin(-), MFG-E8(+), and hCG-R(+), whereas ESC were vimentin(+), cytokeratin(-), MFG-E8(-), and hCG-R(+). Treatment of ESC with MFG-E8 resulted in a 13-fold increase in caspase activity and a 30-fold increase in TUNEL. On the other hand, hCG decreased messenger RNA expression of Bax in ESC. CONCLUSION(S) Milk fat globule-epidermal growth factor 8 has proapoptotic activity, suggesting participation in endometrial remodeling via an epithelial-stromal cell paracrine effect. Conversely, pregnancy levels of hCG has opposite effects on stromal cells.