Routine laser assisted hatching results in significantly increased clincal pregnancies.

PURPOSE: To elucidate the appropriateness of current indications for assisted hatching (AH) in cleavage stage human embryos and to confirm our preliminary findings that only young patients (about 67%) benefit from AH. METHODS: Prior to transfer, 2 of 3 embryos selected for ET were subjected to laser assisted hatching (LAH). Control group consisted of patients matched by similar characteristics and protocol except LAH was not performed. RESULTS: The clinical pregnancy rate in women < or = 36 years was 64.9% (24/37) for embryos subjected to LAH but was significantly lower (p = 0.029) in the control (33.3%; 10/30). The implantation rate in women < or = 36 years in the test group was 38.1% (40/105) that was significantly higher than that of the control group (17.5%, 14/80; p = 0.0039). CONCLUSIONS: LAH is beneficial for women < or = 36 years but not for women > or = 37 years, for embryos with thin zonae (< or = 16 micron) but not with thick zonae (> or = 17 micron), and for those with repeated failures (37-50%).

Formulation of a protein-free medium for human assisted reproduction.

The optimal concentrations of individual amino acids, antioxidants, vitamins, osmolytes and energy sources were determined using a 1-cell Swiss outbred (SO) and or F(1) [(CBAxC57BL/6J)xSO] mouse assay in Earle's balanced salt solution containing bovine serum albumin. Based on the findings of these experiments, a number of media were formulated. Of these, the medium showing optimal embryo development and a significantly higher blastocyst hatching rate was investigated further. A protein-free medium (ART-7) was formulated and assessed using 1-, 2- and 4-cell SO mouse embryos. The generation of viable human embryos in the ART-7 series of media in micro- and ultra micro-droplet culture under oil with and without cumulus co-culture following intracytoplasmic sperm injection (ICSI) was investigated. The quality of sibling day 2 human embryos generated in the ART-7 media series was statistically comparable to or better than control embryos. The ART-7 medium was not toxic to human spermatozoa. Fertilization by conventional IVF and subsequent embryo development was not affected. A clinical trial of ICSI-derived embryos generated in the protein-free medium, with and without cumulus co-culture, has resulted in clinical pregnancies (10 of 20 transfers) of which two have proceeded to term, and the remaining patients are in various stages of pregnancy.

Intracytoplasmic sperm injection improving embryo quality: comparison of the sibling oocytes of non-male-factor couples.

OBJECTIVE: Our objective was to investigate whether the quality of embryos developed after intracytoplasmic sperm injection (ICSI) is better than that of conventional IVF embryos. METHODS: Nine couples who previously achieved a normal rate of fertilization following IVF and four couples whose normal rate of fertilization was expected were involved in this study. The oocytes from those couples were randomly divided into two groups, group A by conventional insemination and group B by ICSI. The fertilization rate and quality of embryos were compared. RESULTS: Normal fertilization was achieved in 61% of the oocytes (83/136) after conventional insemination. In group B, 69% of the oocytes (99/144) achieved normal fertilization, although only 127 metaphase II oocytes were injected using the ICSI technique. More grade A embryos were obtained when the ICSI technique was used for fertilization than by conventional IVF (35.4 and 24.3%, respectively; P = 0.028). CONCLUSIONS: A similar fertilization rate can be achieved by ICSI in comparison with conventional IVF, when male factor is not involved. Embryos after ICSI have an improved quality.