Infertile Donors Research Articles

Polycystic ovary syndrome patients as oocyte donors: the effect of ovarian stimulation protocol on the implantation rate of the recipient

To evaluate the outcome of oocytes donated by women with polycystic ovarian syndrome (PCOS) compared with oocytes donated by women with mechanical infertility. A retrospective study. The outcome of 159 oocyte donation cycles with oocyte donated by PCOS patients were compared with 69 oocyte donation cycles with oocytes donated by patients with mechanical infertility. We compared the stimulation protocols in the donors to assess if the combination of GnRH analogue (GnRH-a), FSH, and hMG has an advantage over FSH and hMG alone with respect to their effect on fertilization and implantation rates in oocyte donation cycles. When treated with GnRH-a, pregnancy rates in PCOS and mechanical infertility donors were higher than those treated with FSH and hMG alone. The comparison between PCOS and mechanical factor oocyte recipients revealed no significant difference in the pregnancy and abortion rates, but the oocytes of patients with PCOS that were exposed to GnRH-a had a significantly higher implantation rate than those not exposed to GnRH-a. Oocytes obtained from PCOS patients had a fertilization potential equal to oocytes obtained from mechanical infertility donors. Furthermore, because the oocytes of patients with PCOS exposed to GnRH-a had a significantly higher implantation rate, a detrimental role of high LH on oocyte quality seems probable. However, because PCOS has a high familial prevalence, some reservations may arise due to a possible propagation of the problem in the next generation of oocyte donation programs.

Sperm surface proteins after capacitation. Expression of vitronectin on the spermatozoan head and laminin on the sperm tail.

Several integrins recognize as ligands proteins containing the Arg-Gly-Asp (RGD) sequence, such as fibronectin, vitronectin, and laminin. It has been previously demonstrated that oligopeptides containing the RGD sequence competitively inhibit both the adhesion of hamster and human sperm to zona-free hamster eggs and their subsequent penetration. In addition, the appearance of fibronectin on the surface of living human spermatozoa after capacitation has been demonstrated. In this work, it is shown that spermatozoa incubated overnight under capacitating conditions, but not fresh spermatozoa, also display the RGD-containing proteins vitronectin and laminin. Whereas the expression of fibronectin does not appear to be localized to any specific region of the sperm surface, laminin is present solely on the sperm tail, and vitronectin was detected mostly as an equatorial band on the sperm head. The percent of capacitated spermatozoa within each ejaculate reacting with antivitronectin antibodies (51% to 94%) was similar to that observed with antifibronectin antibodies (72% to 100%) in a series of fertile donors, and in a series of infertile men (7% to 98% for vitronectin versus 5% to 100% for fibronectin). In contrast, the percent of spermatozoa displaying laminin was lower, ranging from 2% to 42% for fertile donors and from 5% to 34% for infertile donors, and was unrelated to the expression of fibronectin or vitronectin. The time of appearance of both fibronectin and vitronectin when spermatozoa were incubated under capacitating conditions varied for different sperm donors, suggesting a difference in the process of their expression between different men. The specificity of antivitronectin antibody binding to human spermatozoa was demonstrated by competitive inhibition with purified human vitronectin. That there was no immunologic reaction between the antivitronectin antibodies used and fibronectin was demonstrated both by the failure of free fibronectin to inhibit antivitronectin antibody binding to spermatozoa, and by Dot blot analysis. A partial cross-reaction of the polyclonal antivitronectin antibody with fibronectin was shown by Western blot analysis, but this phenomenon was not present when the monoclonal antivitronectin antibody was used. In addition, both fibronectin and vitronectin could be extracted from capacitated spermatozoa solubilized in Chaps buffer, as shown by Dot blot and Western blot analysis. These observations suggest that vitronectin and fibronectin expressed on the surface of capacitated human spermatozoa could act as a ligand for specific receptors on the egg, and play a role in sperm-oolemmal adhesion.

The relationship between alterations in spermatozoal deoxyribonucleic acid, heparin binding sites, and semen quality

To assess the relationship between spermatozoal deoxyribonucleic acid (DNA) and fertilizing potential. Semen samples were examined from nine fertile donors and six donors without a confirmed pregnancy. All samples were in the normal range for count, morphology, and motility. Spermatozoa from these specimens were stained with acridine orange or Feulgen's reagent. The presence of heparin binding sites was determined by counting the number of spermatozoa that bound to heparin-coated agarose beads. Acridine orange staining demonstrated that in the fertile group 42% +/- 2% of the spermatozoa fluoresced green indicating that the DNA was intact, whereas only 25% +/- 3% of the spermatozoa fluoresced green in the nonfertile group (P less than 0.05). Feulgen's staining revealed that more spermatozoa from infertile donors showed a heterogeneous DNA distribution (P less than 0.05). The DNA content of spermatozoa with heterogeneous distribution of DNA was reduced by 10% compared with those with homogeneous DNA (P less than 0.05). Normal spermatozoa as well as those with DNA anomalies possessed heparin binding sites. These data demonstrated that in donor specimens with normal counts, morphology, and motility, a higher percentage of spermatozoa possess less and/or denatured DNA in the infertile group compared with the fertile donors. In contrast, the surface membranes of spermatozoa with altered DNA have heparin binding sites as do spermatozoa with intact DNA.

A comparison of reproductive performance of fertile versus infertile donor mares

A comparison of reproductive performance of fertile versus infertile donor mares

Analysis of Human Spermatozoal Fertilizing Ability Using Zona-Free Ova

An in vitro fertilization assay employing zona-free hamster eggs was used to analyze human spermatozoal fertilizing ability. Human spermatozoa were preincubated for 18 to 20 hours in Biggers, Whitten, and Whittingham's medium (1971) at a concentration of 1 X 10(7) sperm/ml prior to the addition of zona-free superovulated hamster eggs. Eggs were examined microscopically 2 hours later for evidence of swelling or decondensing sperm heads in the cytoplasm. A total of 6266 eggs were examined in assays for both suspected fertile and infertile donors; 50 eggs/sample were examined. The percentage fertilization was found to range from 14% to 100% in the suspected fertile group with an average of 56.3%. The sperm concentration in this fertile group ranged from 22 to 303 million/ml with an average of 114. The suspected infertile samples yielded fertilization rates of 10% or less and an average count of 50.6 million/ml. These data suggest that human spermatozoa fuse with the vitelline membrane of zona-free hamster eggs and decondense with varying efficiencies. The percentage of fertilization in this cross-species system did not show a significant correlation with sperm concentration or motility. However, suspected infertile samples always yielded 10% or less fertilization in this assay. This method may have potential value as a diagnostic tool in evaluating human spermatozoal fertilizing capacity which avoids the ethical and logistcal problems associated with fertilization of human eggs in vitro.

Embryo transfer in fertile and infertile cows

Pregnancy rates were compared between fertile and infertile donors in an embryo transfer program. Non-surgical embryo transfer techniques were utilized for examination of uterine contents and collection of ova. Recovered embryos were transferred surgically into suitable recipients. Three groups of donors were included in this investigation: a) fertile; b) infertile, due to known causes (diagnosed); c) infertile, due to unknown causes (undiagnosed). There were 11.3, 6.6 and 8.0 corpora lutea; 6.8, 1.2 and 1.0 fertilized ova; 3.6, 0.8 and 0.4 pregnancies per superovulation for the fertile, diagnosed and undiagnosed groups, respectively. In 23 months, unsuperovulated, fertile, diagnosed and undiagnosed donors yielded 17, 15 and 9 fertilized ova which resulted in 15, 13, and 3 pregnancies, respectively. Donors treated for uterine infections, adhesions or cystic ovaries prior to superovulation responded with 10.1, 7.6 and 4.1 corpora lutea; 1.9, 4.0 and 0.3 fertilized ova; 1.1, 3.3 and 0.2 pregnancies per superovulation, respectively. Relatively few viable embryos were recovered from donors with chronic cystic ovaries or from infertile cows of unknown etiology. Infertile donors, when compared to fertile donors, were unproductive when used for embryo transfer.