M. Dirnfeld

A prospective randomized controlled study of the effect of short coincubation of gametes during insemination on zona pellucida thickness

A prospective, randomized study was conducted to evaluate the thickness of zona pellucida (ZP) after brief or standard exposure of human oocytes to spermatozoa, and to determine the correlation between ZP thickness, fertilization rate and embryo quality. The mean ZP thickness 48 h after insemination was found to be significantly less in fertilized oocytes than in non-fertilized oocytes in all treated groups (13.72 ± 3.0 µm and 15.08 ± 2.5 µm, respectively; p < 0.007). Zona pellucida thickness correlated positively with embryo quality. Brief exposure of gametes was found to influence ZP thickness. The ZP was significantly thinner after brief and intracytoplasmic sperm injection (ICSI) exposure of oocytes to spermatozoa than after standard in vitro fertilization (IVF). The mean ZP thickness 24 and 48 h after fertilization was significantly greater in standard IVF (16.43 ± 2.8 µm and 15.22 ± 2.7 µm, respectively) than in either the brief exposure or ICSI groups (12.78 ± 2.4 µm and 13.01 ± 3.5 µm vs. 13.46 ± 2.2 µm and 13.16 ± 2.4 µm; p < 0.0001).

Oxidative parameters of embryo culture media may predict treatment outcome in in vitro fertilization: a novel applicable tool for improving embryo selection

OBJECTIVE To examine whether the oxidative status of an individual embryo before transfer may predict chances of implantation. DESIGN A prospective laboratory study. SETTING An IVF unit in a university-affiliated hospital. PATIENT(S) One hundred thirty-three women undergoing IVF-ET treatment cycles. INTERVENTION(S) Before ET, 10 μL of embryo culture medium was retrieved individually from each embryo and the oxidative status assessed by the thermochemiluminescence (TCL) analyzer. MAIN OUTCOME MEASURE(S) The occurrence of pregnancy. Two parameters were recorded: the TCL amplitude after 50 seconds (H1) and the TCL ratio. These were compared with demographic, clinical, and laboratory parameters and treatment outcome. All data underwent statistical analysis. RESULT(S) Altogether 284 embryos were transferred in 133 ET cycles. Forty-one pregnancies occurred (31%). For embryos transferred after 72 hours (77 transfers), the highest H1 levels in each group of transferred embryos correlated with the occurrence of pregnancy. The combination of maximal intracohort H1 level <210 counts per second with a TCL ratio of ≤ 80% had a positive predictive value of 70.6% for the occurrence of pregnancy. CONCLUSION(S) The oxidative status of the early embryo in IVF is associated with the chances of implantation. Assessment of the oxidative status of embryos in culture media before transfer may serve as an applicable tool for improving embryo selection in light of the legal limitations of the number of transferred embryos allowed.

p27 and its ubiquitin ligase Skp2 expression in endometrium of IVF patients with repeated hormonal stimulation

This preliminary study examined a possible effect of long duration repeated hormonal stimulation on the endometrium using a molecular tool. The expression of the hormone stimulated, cell cycle regulators, p27 and its ligase S-phase kinase-interacting protein2 (Skp2), were assessed in 46 endometrial samples of patients who underwent repeated IVF cycles (3-21). Skp2 protein is usually undetectable in normal tissue and can be demonstrated only in rapidly dividing cells. Samples from non-stimulated, normal cycling women served as control group A. Samples of endometrial carcinoma served as control group B. In secretory endometrium, the expression of p27 was found to be lower and Skp2 higher in the study group compared with control group A. Moreover, in 25% of patients of the study group, Skp2 expression was significantly higher (P < 0.05) compared with control group A, reaching concentrations demonstrated in endometrial carcinoma. The findings of this study suggest that repeated hormone stimulation cycles may disrupt endometrial physiology, potentially towards abnormal proliferation. These changes in protein expression are described for the first time in IVF patients and should be further investigated.

Live Birth from the Transfer of a Severely Fragmented Embryo Observed by Morphokinetics

We report a live birth from a heavily fragmented embryo which continued cleavage to a fully expanded blastocyst. A 32-year-old patient underwent 2 IVF cycles without achieving pregnancy. In the first cycle, 2 embryos with fragmentation were transferred; in the second, all embryos were fragmented and no embryo transfer was performed. In a third cycle, 12 oocytes were retrieved and 11 of them were fertilized. On day 2, all 11 embryos started to unwind to fragments. By careful annotation, using the time-lapse EmbryoScope, we observed that one embryo continued division as expected, discarding all fragments aside. On day 5, this embryo showed promising annotation according to our lab model. The embryo was transferred into the uterus and resulted in the birth of a healthy baby at term. To our knowledge, this is the first case report assisted by EmbryoScope where a healthy baby was delivered from a fragmented embryo.