Aaron L. Wilcox

Elevated sperm chromosome aneuploidy and apoptosis in patients with unexplained recurrent pregnancy loss.

OBJECTIVE To evaluate sperm chromosome aneuploidy and semen quality in 24 partners of women with unexplained recurrent pregnancy loss and to analyze the data in relation to sperm apoptosis data. METHODS Semen quality parameters and sperm chromosome aneuploidy for chromosomes X, Y, 13, 18, and 21 were evaluated in the recurrent pregnancy loss patients, fertile controls, and a control group of men from the general population. RESULTS The mean aneuploidy rate in the recurrent pregnancy loss group was 2.77 ± 0.22, significantly higher (P < .005) than in either the general population (1.48 ± 0.12) or in fertile (1.19 ± 0.11) control groups. In the recurrent pregnancy loss patients, the percentage of aneuploid sperm was correlated to the percentage of apoptotic sperm (r = .62, P < .001). Normal morphology was diminished in the patient group, compared with the general population group (P < .01) and the donor group (P < .001). CONCLUSION These data indicate that some recurrent pregnancy loss patients have a significant increase of sperm chromosome aneuploidy, apoptosis, and abnormal sperm morphology. This study demonstrates a new possible cause of recurrent pregnancy loss.

Chromosome 15 aneuploidy in the sperm and conceptus of a sibling with variable familial expression of round-headed sperm syndrome.

OBJECTIVE To characterize rates of chromosome aneuploidy in sperm from three siblings, one of whom had an IVF/ICSI conceptus with trisomy 15. DESIGN Blind evaluation of the sperm chromosome aneuploidy rates, semen quality, and sperm ultrastructure. SETTING IVF clinic and university-based andrology research laboratory. PATIENT(S) Three brothers, two of whom underwent infertility evaluation and therapy. MAIN OUTCOME MEASURE(S) Semen from three siblings was coded and blindly evaluated for standard World Health Organization semen quality variables and sperm ultrastructure. Sperm were decondensed and hybridized with fluorescent probes specific for chromosomes X, Y, 13, 15, 18, and 21, then evaluated microscopically to determine the aneuploidy rate for those chromosomes. RESULT(S) Two siblings had increased round-headed morphology on standard morphology evaluation, which was confirmed using electron microscopy. The sperm aneuploidy rate was significantly increased for chromosome 15 in sibling 1, the father of a conceptus with trisomy 15. Aneuploidy rates were also slightly increased for chromosomes X, Y, and 18 in sibling 1. CONCLUSION(S) This is the second report of increased sperm chromosome aneuploidy in infertile patients with round-headed sperm. Although ICSI is successful in treating this syndrome, the risk for aneuploidy of the conceptus may be increased. Other studies have reported an increased incidence of sperm chromosome aneuploidy in some infertile patients, but this is the first report of aneuploidy in both the sperm and conceptus of a patient undergoing IVF/ICSI.

Development of In Vitro-Matured Oocytes from Porcine Preantral Follicles Following Intracytoplasmic Sperm Injection

Abstract The objective of this study was to assess fertilization and embryonic development following intracytoplasmic sperm injection (ICSI) of oocytes from porcine preantral follicles matured in vitro. Also, another aim was to describe actin filament distribution during fertilization and embryonic development of those oocytes after ICSI as one of the factors assessed. Preantral follicles isolated from prepubertal porcine ovaries were cultured in a system that supports follicular development. After in vitro maturation, the oocytes were fertilized by ICSI or conventional fertilization in vitro (IVF). Actin filaments of the fertilized oocytes and embryos produced by ICSI or IVF were stained by rhodamine-phalloidin and visualized by fluorescence microscopy. ICSI resulted in 64% fertilization of porcine preantral follicle oocytes matured in vitro. Of those, 51% of the fertilized oocytes cleaved and 21% developed to the blastocyst stage. No significant differences in percentages of oocyte fertilization, cleavage, and blastocyst formation were observed between ICSI and IVF (53%, 45% and 16%, respectively). Actin filament distribution during fertilization and embryonic development of ICSI- or IVF-fertilized oocytes from porcine preantral follicles was similar to that of oocytes derived from antral follicles and fertilized by standard IVF. These results indicate that oocytes from porcine preantral follicles matured in vitro following ICSI can undergo fertilization and subsequent embryonic development.

Sperm chromosome aneuploidy as related to male factor infertility and some ultrastructure defects.

Some men have elevated levels of sperm chromosome aneuploidy. In this study, we have evaluated and summarized sperm aneuploidy rates in male infertility patients and control groups. The mean aneuploidy rate for five chromosomes (X, Y, 13, 18, 21) was 1.2 ± 0.1 for fertile controls, 1.4 ± 0.1 for a general population control group, and 5.8 ± 1.14 for the patients. When the patients were classified by the type of male factor infertility, the total aneuploidy rate was 2.6 ± 0.3 in men with moderately diminished semen quality (n = 7), 4.0 ± 0.3 patients with severe teratoasthenooligozoospermia, and 15.9 ± 3.8 for men with rare ultrastructure defects such as round head only syndrome or severe tail agenesis. Some infertility patients have a severely elevated level of sperm chromosome aneuploidy, which may contribute to infertility or diminish the likelihood of a successful outcome from IVF/ICSI. The severity of sperm chromosome aneuploidy appears to be proportional to the severity of abnormal semen quality: in particular, abnormal morphology. The high rates of aneuploidy in patients with severe ultrastructure defects suggest that caution should be employed in counseling those patients prior to IVF/ICSI.

Comparison of four media types during 3-day human IVF embryo culture

The aim of this study was to compare the effectiveness of human tubal fluid (HTF), G1.2, Sage Cleavage and Life Global media for IVF outcome during 3-day culture of human embryos. A three-phase auto-controlled study was conducted in which IVF outcome was compared between (1) HTF and G1.2, (2) HTF and Cleavage, and (3) Cleavage and Life Global. In phase 1, no differences in embryo quality were observed between HTF and G1.2. However, embryos derived from intracytoplasmic sperm injection (ICSI) displayed significantly improved quality when grown in HTF versus G1.2. No differences in pregnancy and implantation rates were observed in cases where embryos transferred were grown exclusively in HTF or G1.2 media. In phase 2, embryo quality was significantly improved for embryos cultured in Cleavage versus HTF media (P < 0.001). However, pregnancy, implantation and spontaneous abortion rates were similar between the two media. In phase 3, there were no differences in embryo quality, pregnancy, implantation, and spontaneous abortion rates between Cleavage and Life Global media. Overall, the data indicate that Life Global and Cleavage media yield similar results in a 3-day IVF culture programme. Cleavage medium is superior to HTF, as evidenced by significantly improved embryo quality (P < 0.001). Meanwhile, HTF medium is superior to G1.2 for ICSI cases.

Comparison of 5% and ambient oxygen during days 3-5 of in vitro culture of human embryos

OBJECTIVE To compare the effect of two oxygen concentrations used during days 3-5 of human embryo culture on embryo quality and pregnancy outcome. DESIGN Retrospective analysis of the use of two culture conditions. SETTING University-based infertility clinic. PATIENT(S) Three hundred eighty-two patients undergoing IVF. INTERVENTION(S) Embryos were cultured in 5% CO(2) balanced ( approximately 20% O(2)) gas phase until day 3 then assigned to approximately 20% or reduced (5%) oxygen concentration groups and cultured until ET. MAIN OUTCOME MEASURE(S) Embryo quality, pregnancy rates, and implantation rates. RESULT(S) There were no differences in demographic features (age, type of infertility) between the two groups. The embryo scores at day 3 and day 5, blastulation rate, and transfer score did not differ between groups. No differences were observed between the 5% and 20% oxygen concentrations in the chemical pregnancy rate (71.27% vs. 78.72%), clinical pregnancy rate (58.56% vs. 64.36%), or implantation rate (44.06% vs. 44.16%). CONCLUSION(S) Reduced oxygen concentration in the gas mixture from day 3 until ET did not support better embryo development or result in higher pregnancy or implantation rates. These data do not support the hypothesis that beneficial effects of reduced oxygen concentration can be gained by employing the strategy during the latter stages of embryo culture (days 3-5) only and highlight the need for further studies through all stages of in vitro culture.

Intracytoplasmic Morphology-Selected Sperm Injection

Approximately 40% of sterility in couples can be attributed to male subfertility and intracytoplasmic sperm injection (ICSI) has become a powerful tool in assisted reproduction to overcome male infertility. Intracytoplasmic morphologically selected sperm injection (IMSI) is an advanced and sophisticated method of ICSI, where prior to sperm injection the morphology of the sperm is evaluated under high magnification. In addition, the IMSI procedure involves a few minor modifications in sperm preparation which are not carried out during the conventional ICSI procedure, such as the use of MSOME criteria, the requirement for a glass-bottomed dish for selection, prolonged sperm manipulation following separation from the seminal fluid, and sperm storage prior to microinjection. These variations are discussed in this chapter.