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The aim of this study was to evaluate the information and the factors that contribute to the decision to accept and choose single embryo transfer (SET) in females and males. Fifty-four females and males undergoing SET were interviewed separately using a structured questionnaire. The women were significantly more satisfied with the information than the men (odds ratio 3.3), but the decision to accept SET was nevertheless more difficult for women (OR 3.1). Only one-third of both female and males were aware of the increased maternal risks with twin pregnancies. There was a tendency that the women who accepted SET had previous children, shorter duration of infertility, and were younger. Cryopreservation of embryos and a good pregnancy chance were important irrespective of gender. The female needs more support to choose SET. The male needs better information and further involvement in decision-making. The females were more aware of the fetal risks, but the awareness of the increased maternal risks with twin pregnancies was low.

Chromosome preparations were made from 25 cleaved abnormal human embryos at the two-to eight-cell stage after in vitro fertilization. Morphologically, these embryos showed either variable degrees of degeneration or an abnormal number of pronuclei before first cleavage. Among 14 successfully karyotyped embryos, only 3 had a normal chromosomal complement. Eleven showed chromosomal abnormalities, including triploidy, hapioidy, and mosaicism. This finding documents a high incldence of chromosomal errors in morphologically abnormal early preimplantation embryos.

To evaluate the relationship between sperm nuclear vacuoles and sperm morphology and to investigate the influence of the rate of spermatozoa with head vacuolization (SVR) in a seminal sample on the clinical outcomes in couples undergoing intracytoplasmic sperm injection. 26 patients undergoing infertility investigations were included and were divided in two groups according to an SVR ≤ 20,28 % (Group A) or > 20,28 % (Group B), and were investigated to verify the influence of SVR on the fertilization rate, embryo quality, pregnancy and implantation rates. Abnormal spermatozoa with nuclear vacuoles were significantly higher (p < 0.001) than the percentage of normal spermatozoa with nuclear vacuoles. Patients in group A had a percentage of abnormal sperm with nuclear vacuole significantly lower compared to group B (p < 0,001), but there was no difference in the percentage of normal sperm with nuclear vacuoles. Fertilization rates and the number of top quality embryos did not differ between the two groups. The pregnancy and implantation rates were significantly higher in Group A compared to Group B (respectively p < 0,05 and p < 0.001). For the first time, we propose a cut off value in the proportion of sperms with nuclear vacuolization on the total of sperm in seminal samples, and demonstrate a relationship between SNV and clinical outcomes after ICSI. The SNV rate could be introduced as an easy diagnostic evaluation prior to perform an ICSI cycle.

To describe a new technique for freezing individually isolated spermatazoa from testicular biopsies, epididymal aspirates and oligospermic semen samples Samples were evaluated for the presence of motile sperm before cryopreservation. Motile or twitching sperm were isolated with an ICSI needle for single sperm cryopreservation. Selected sperm were loaded on the High Security Straw (HSV; Irvine Scientific; Irvine,CA), in ~0.5 µl of fluid to facilitate recovery. The sample was also frozen using conventional methodology in cryovials (100–1000 µl aliquots). In both freezing techniques, the samples were slow cooled. Test-yolk buffer-glycerol (Irvine) was used as the cryoprotectant. Test-thaws were performed to assess sperm recovery and motility. Six men with azoospermia had single sperm cryopreservation, as well as freezing aliquots of their testicular or epididymal sperm in traditional cryovials. In addition, two men with oligospermia also had individual sperm selected and frozen. In all 8 cases, the ~0.5 µl of fluid containing sperm was quite easily unloaded from the HSV straw during thawing. The percent sperm recovery ranged from 33% to 100%. Motility was evident in all but one sample. In six cases, the sperm were used for intracytoplasmic sperm injection of mature oocytes. Fertilization occurred in all but one case. In this study, we report the first clinical pregnancy with this technique. This pregnancy was remarkable in that a single motile sperm identified and selected in the initial testicular preparation was successfully frozen. We were able to subsequently recover this sperm, fertilize an oocyte and the resultant embryo gave rise to a live birth. The methodology described in this preliminary report offers a new modality for sequestering small numbers of sperm. It may be particularly useful in cases involving severe impairment of spermatogenesis, where extensive screening may be necessary to find a few viable sperm.

To establish a freeze-thawing method for unfertilized oocytes with a high success rate, we examined several conditions for freeze-thawing. The effects of EDTA and cocultures in oviducts on the development of embryos fertilized in vitro after thawing were also studied. In the first experiment, unfertilized oocytes that were frozen in 1.5 Mdimethylsulfoxide (DMSO) supplemented with 0.2 Msucrose by a slow freeze-thawing method showed the best results (fertilization rate, 71.9%; blastocyst rate per frozen oocyte, 18.8%). The proportion of embryos that developed to blastocysts was significantly higher when DMSO was added at 4‡C than at room temperature (39.4 vs 19.4%; P<0.01). The addition of EDTA (10 ΜM)to the culture medium did not promote embryo development after fertilization in vitro. However, the rate of development of in vitro fertilized embryos to blastocysts after thawing was significantly higher when the embryos were cultured in oviducts in vitro than the rates in control cultures and those cultured with EDTA (blastocyst rate from fertilized oocytes, 71.4 vs 51.0 and 52.8%, respectively; P<0.01). Unfertilized mouse oocytes can be cryopreserved successfully by a slow freeze-thawing method with the addition of 1.5 MDMSO and 0.2 Msucrose at low temperatures, and coculture with oviducts enhances the development of embryos that are fertilized in vitro after thawing.