Pregnancy after embryo biopsy and coamplification of DNA from X and Y chromosomes.
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Biopsy↗
Biomedical subjects
Z Rosenwaks
Publications and source records attributed to Z Rosenwaks.
Controversies and opinions in clinical microsurgical fertilization.
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Evaluation Studies as Topic↗
Relation between antibodies to Chlamydia trachomatis and spontaneous abortion following in vitro fertilization.
BACKGROUND: Many couples undergo in vitro fertilization due to occlusion of the fallopian tubes. Chlamydia trachomatis infections are a major cause of this tubal damage. Since this organism has also been associated with poor pregnancy outcome, we investigated whether a past exposure to C. trachomatis was associated with spontaneous abortion following in vitro fertilization and embryo transfer. METHODS: Sera from 145 women undergoing IVF were diluted 1:128 and tested for IgG antibodies to C. trachomatis by an immunoperoxidase assay, using infected cells fixed to slides. All subjects and their partners were negative for C. trachomatis by culture or by DNA hybridization. RESULTS: Serological evidence of a past chlamydial infection was observed in 33.8% of the women. The incidence of antichlamydial IgG was greater (P less than 0.001) in women whose infertility was due to known tubal disease (37 of 78; 47.4%) than in women whose infertility was due to other causes (12 of 67; 17.9%). Spontaneous abortions after embryo transfer occurred in 20% of the subjects. The incidence of antichlamydial IgG in aborting women (20 of 29; 69.0%) was greater (P less than 0.001) than the incidence in either women with successful pregnancies (9 of 38; 23.7%) or women who did not become pregnant (20 of 78; 25.6%) after IVF. No relation was observed between antichlamydial antibody status and maternal age, the number of oocytes aspirated, the number of oocytes fertilized, and the number of embryos transferred. CONCLUSIONS: A previous infection with C. trachomatis may increase susceptibility to subsequent spontaneous abortion, even in the absence of a detectable current infection.
Abortion, Spontaneous↗
Improved pregnancy outcome with gonadotropin releasing hormone agonist (GnRH-a) stimulation is due to the improvement in oocyte quantity rather than quality.
The impact of gonadotropin releasing hormone agonist (GnRH-a) on the quality and quantity of oocytes harvested in in vitro fertilization-embryo transfer (IVF-ET) patients was studied by comparing the results for patients stimulated with gonadotropin alone and with gonadotropin plus GnRH-a. Adding GnRH-a significantly improved the viable pregnancies per transfer and reduced the spontaneous abortions, which seemed to improve oocyte quality. However, when oocyte quality was evaluated by the fertilization rate and the implantation and delivery rates per embryos transferred, there were no significant difference in the results, indicating that GnRH-a did not improve the oocyte quality. On the other hand, GnRH-a significantly increased the average number of oocytes harvested, fertilized, and transferred, and this increased number of oocytes transferred has been demonstrated to increase pregnancy and multiple-pregnancy rates. Multiple pregnancy with more embryos implanted would significantly reduce the abortion rate. Abortion rates decreased inversely to the number of embryos implanted. Our data strongly suggest that the efficacy of GnRH-a on IVF-ET patients was due more to the quantity increase than the quality of embryos transferred.
Embryo Implantation↗
Relation between antisperm antibodies and the rate of fertilization of human oocytes in vitro.
To clarify further the role of antisperm antibodies in in vitro fertilization, the occurrence of antisperm antibodies on ejaculated sperm and in sera was determined by the immunobead binding assay in 67 couples after an unsuccessful in vitro fertilization cycle. Antisperm antibodies in maternal sera were associated with a failure of oocyte fertilization (P less than 0.02) or with fertilization of only 9-19% of the oocytes (P less than 0.01) in vitro. Antisperm antibodies were detected in sera from 13 of 24 women (54.2%) where no fertilization occurred, 9 of 14 women (64.3%) where less than 20% of the oocytes fertilized, and 3 of 19 women (15.8%) where greater than 40% of the oocytes fertilized. Antisperm antibodies in these sera were mostly IgG and directed against the sperm tail. Antibodies on the surface of ejaculated motile sperm were also associated with a low (9-19%) fertilization rate (P less than 0.01). Sperm-bound antibodies were detected in 2 of 24 men (8.3%) where no fertilization occurred, 5 of 14 men (35.7%) where less than 20% of the oocytes fertilized, and 0 of 19 men where fertilization was greater than 40%. Sperm-bound antibodies were mainly IgA and were tail-directed. Antisperm antibodies in sera of males were not related to the rate of fertilization.(ABSTRACT TRUNCATED AT 250 WORDS)
Adult↗
Evaluation of Vero cell co-culture system for mouse embryos in various media.
Our objective was to evaluate and compare the efficacy and mechanisms of co-culturing mouse embryos with Vero cells in both Dulbecco's modified Eagle's medium/Ham's F12 (DMEM/F12) and human tubal fluid (HTF) culture medium. Two-cell CB6F1 mouse embryos were cultured either in the presence of Vero cells (group A) or in culture medium alone (group B). In DMEM/F12 significantly more morulae developed in group A than in group B on day 3 (91 versus 23%; P less than 0.01). In contrast, the mouse embryos grew rapidly in HTF and significant differences were noted only in later embryonic stages (on day 5; 86% and 50%, P less than 0.01 of group A and B respectively, hatching or hatched). Similar experiments using DF1 and ICR mouse strains also revealed enhanced embryo development in the presence of Vero cells. To determine whether the embryo-enhancing effects of Vero cells were due to the removal of toxins or to the secretion of embryotrophic factors, ICR mouse embryos were cultured in fresh media with cells (group A), without cells (group B) and in cell-free conditions using cell-conditioned media which were obtained in the presence (group C) or absence (group D) of embryos. These results demonstrated that completion of hatching was highest (52%; P less than 0.01) in group A after 6 days in culture. There were no significant differences between groups B, C and D (rates of total hatching 18, 17 and 17%, respectively). It is concluded that Vero cells improve the development of mouse embryos and this is likely to be due to removal of substances inhibitory to development.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Implantation enhancement by selective assisted hatching using zona drilling of human embryos with poor prognosis.
Assisted hatching by zona drilling using acidic Tyrode's solution was performed during three randomized trials in 330 in-vitro fertilization patients. The trials were designed in order to study the overall effect of the procedure and whether characteristic patient [i.e. maternal age and basal levels of follicle stimulating hormone (FSH)] and embryonic features (i.e. zona pellucida thickness) are important for the decision to perform assisted hatching routinely. Couples (n = 137; Trial 1) in whom the female partner had normal basal FSH levels were randomized in a control group (without micromanipulation) and a zona drilling group (all embryos micromanipulated). The incidence of implantation (67/239; 28%) of zona-drilled embryos compared favourably with that of control embryos (49/229; 21%), but the difference was not significant. Retrospective analysis revealed that those embryos whose zonae were thicker than 15 microns were rescued. In order to test the validity of this finding, selective assisted hatching was performed on embryos with a poor prognosis in 163 other patients (Trial 2). The couples were randomized into a control group and a group in which embryos were selectively zona-drilled, based on zona thickness and other embryonic features. The rate of embryonic implantation in the selectively zona-drilled group was 25% (70/278), significantly (P less than 0.05) higher than that of control embryos (51/285; 18%). Although it was demonstrated retrospectively and prospectively that assisted hatching by zona drilling is effective in embryos with thick zonae (greater than or equal to 15 microns), patients whose embryos have thin (less than 13 microns) zonae may be jeopardized by the procedure.(ABSTRACT TRUNCATED AT 250 WORDS)
Embryo Implantation↗
Current status of in vitro fertilization and the new reproductive technologies.
The rapid pace of development and refinement of in vitro fertilization and ancillary assisted reproductive techniques warrants periodic review to keep the reader abreast of the growth in this clinical and scientific arena. Although not exhaustive, this review targets key areas of special interest and significance, including oocyte and embryo micromanipulation for assisted fertilization and genetic analysis, advances in the approach to ovarian stimulation, and examination of the scientific model afforded by ovum donation.
Female↗
Early pregnancy wastage in IVF (in vitro fertilization) patients.
Human in vitro fertilization is characterized by a low efficiency of implantation. Possible mechanisms for pregnancy loss are discussed. Embryo viability or quality, abnormal implantation, and delayed or absent corpus luteum rescue may all play a role in pregnancy wastage. Defining the possible mechanism for these losses may allow hormonal treatment to correct specific abnormalities.
Abortion, Spontaneous↗
Tumor necrosis factor is present in maternal sera and embryo culture fluids during in vitro fertilization.
The incidence of tumor necrosis factor alpha (TNF) in sera of women undergoing in vitro fertilization (IVF) and in embryo culture fluids was evaluated using an enzyme-linked immunosorbent assay. Just prior to embryo transfer to the uterus, 24 of 49 maternal sera (49.0%) contained TNF. The incidence and range of TNF concentrations (84-920 pg/ml) did not differ between women with eventual successful pregnancies and women who subsequently suffered preclinical or clinical abortions. At 8 days post-embryo transfer, 24 of 56 sera (42.9%) contained TNF. Again, the occurrence of TNF was of no predictive value for the eventual outcome of the pregnancy. TNF was also detected in culture fluids from the in vitro fertilized eggs of 12 of 49 women (24.5%). In 9 women, TNF was detected in each of the embryo culture fluids tested. As was the case for sera, the presence or absence of TNF in the culture fluids was unrelated to pregnancy outcome. In 21 patients, paired sera and culture fluids were analyzed. In 9 of 10 women with TNF in their culture fluids, TNF was also present in the corresponding serum. Of 12 women with serum TNF, 9 also had TNF in their culture fluids. Since the culture fluids contained 10% maternal sera, it appeared that in most cases TNF in the culture fluids was derived, at least in part, from the serum. However, in 6 of 9 women TNF levels in the cultures exceeded levels in the corresponding sera. TNF was also identified in 2 of 10 culture fluids in which Plasmanate was substituted for serum.(ABSTRACT TRUNCATED AT 250 WORDS)
Culture Media↗
The window of embryo transfer and the efficiency of human conception in vitro.
Women with ovarian failure transferred with donated oocytes provide a unique in vivo model for the elucidation of the window of implantation and efficiency of reproduction in the human. Throughout 52 ovum donation cycles, the temporal window of endometrial receptivity was tested by replacing 2- to 12-cell embryos between days 16 and 24 of hormonally and histologically defined cycles. Of 37 transfers within days 17 to 19, 15 (40.5%) conceptions occurred. Twelve (32.4%) have reached viability. Of 11 patients transferred on days greater than or equal to 20, none conceived. Likewise, no pregnancies were achieved with 4 transfers on cycle day 16. Analysis of multiple embryo transfers within the suggested window of endometrial receptivity (days 17 to 19) revealed 14 of 24 (58.3%) to be conception cycles. considering only transfers with two or more embryos, at least one of which is of high quality (grades 1 to 2), yielded a 63.2% pregnancy rate. The results indicate a very high efficiency for in vitro fecundity provided optimal conditions are attained. The concepts leading to success in the ovum donation model should set the course for continued research toward improving results in other forms of assisted reproduction.
Abortion, Spontaneous↗
Gonadotropin-releasing hormone agonist-induced ovarian hyperstimulation: low-dose side effects in women and monkeys.
OBJECTIVE: To determine whether low (subtherapeutic) doses of gonadotropin-releasing hormone agonists (GnRH-a) can cause ovarian hyperstimulation. DESIGN: The study is in two parts: a preliminary clinical trial of women and a follow-up study in laboratory primates. SETTING: Normal human volunteers were studied in an academic research environment; primates were in a conventional laboratory setting. PATIENTS, PARTICIPANTS: Human volunteers were selected on the basis of apparent normal health. The monkeys were believed to be of normal reproductive status. INTERVENTIONS: Gonadotropin-releasing hormone agonists were administered at subtherapeutic doses. MAIN OUTCOME MEASURES: After observing ovarian hyperstimulation in two of five women receiving low doses of GnRH-a, a study was specifically designed to test the hypothesis that at low (subtherapeutic) doses of GnRH-a the "flare-effect" can be sustained without achieving down regulation. RESULTS: The data in women and monkeys suggest that a highly individualized response to low GnRH-a doses can be manifested as ovarian hyperstimulation. CONCLUSION: Four points of interpretation are offered: (1) that subtherapeutic doses of GnRH-a can cause ovarian hyperstimulation and related sequelae; (2) this may be a unique observation in that, typically, lower doses of medications have a lower incidence of negative side effects; (3) the findings suggest that GnRH-a prescribed in self-administration regimens may be more prone to such problems in noncompliant patients; and (4) the hyperstimulation response of the ovaries to low GnRH-a doses may indicate a new approach to controlled ovulation induction, although wide individualism was found.
Administration, Intranasal↗
Late luteal estradiol patterns are a better prognosticator of pregnancy outcome than serial beta-human chorionic gonadotropin concentrations.
OBJECTIVE: Since the corpus luteum (CL) is known to play an important role in early pregnancy, its activity could possibly be a marker for pregnancy outcome. DESIGN: The late estradiol (E2) concentration in 48 viable pregnancies and 39 pregnancies which resulted in spontaneous abortions after in vitro fertilization and embryo transfer were used to evaluate such predictability. SETTING: All patients studied were of the Center for Reproductive Medicine at Cornell University Medical College. PATIENTS, PARTICIPANTS: Eighty-seven patients. INTERVENTIONS: None. MAIN OUTCOME MEASURE: Serum E2 and human chorionic gonadotropin (hCG) concentrations on day +11, +13, +15 (day +1 = day of ovum pick-up) were measured and studied. RESULTS: The late luteal CL activity after rescue had a positive correlation with the number and quality of the implanted embryos. Reduced CL activity was indicative of abortion. The late luteal E2 pattern when compared with hCG doubling time had a better abortion predictability (37.8% versus 63.9%, respectively). CONCLUSION: Corpus luteum activity demonstrated to be a better prognosticator of abortion than serial beta-hCG titers.
Chorionic Gonadotropin↗
A human in vivo model for the luteoplacental shift.
OBJECTIVE: To study the onset of early placental steroidogenesis in humans. DESIGN: Ovarian failure patients who were administered a constant dosage of exogenous estradiol (E2) and progesterone (P) during the luteal phase and early gestation and who conceived after transfer of donated oocytes were used to assess serum hormonal changes throughout early pregnancy. SETTING: Hormonally replaced ovarian failure patients undergoing an egg donation program in an academic research environment. PATIENTS, PARTICIPANTS: Nine human volunteers desiring egg donation. INTERVENTIONS: Replacement of E2 and P. MAIN OUTCOME MEASURE: Serum E2 and P concentrations were measured weekly throughout the patients' gestations. RESULTS: Serum E2 and P levels of these patients remained low in early gestation but significantly increased 25 +/- 1.6 and 34 +/- 1.6 days post embryo transfer, respectively. Regression lines of E2 (r = 0.99; P less than 0.01) and P (r = 0.95; P less than 0.005), intersected basal hormonal levels approximately 3 weeks after transfer. CONCLUSION: This estimate of the onset of placental steroidogenesis is equal to the 5th gestational week as calculated by the patients' last menses, approximately 3 weeks earlier than previously reported.
Corpus Luteum↗
Partial zona dissection or subzonal sperm insertion: microsurgical fertilization alternatives based on evaluation of sperm and embryo morphology.
OBJECTIVE: To establish guidelines for application of partial zona dissection, subzonal sperm insertion, and regular in vitro fertilization (IVF) in severe male factor patients. DESIGN: Two studies were performed: partial zona dissection and IVF was applied in 57 couples during the first period, and subzonal sperm insertion was also applied in a second group of 47 couples. SETTING: Procedures were performed in an academic research environment. PATIENTS, PARTICIPANTS: Couples who failed fertilization previously, others not acceptable for IVF, and a third group in whom IVF was expected to fail. INTERVENTIONS: Oocytes were micromanipulated with either partial zona dissection or subzonal sperm insertion, or the zona pellucida was left intact. Embryos were replaced in patients prophylactically treated with methylprednisolone and antibiotics. MAIN OUTCOME MEASURES: Because several microsurgical fertilization techniques are now available, this study was performed to compare sperm parameters, embryo morphology, fertilization, and implantation rates after application of two successful micromanipulation procedures. RESULTS: Twenty-one pregnancies were established in 104 patients, 5 definitely from subzonal sperm insertion and 4 from partial zona dissection. Patients who failed IVF before had a similar chance of pregnancy after the use of micromanipulation, as first time patients (9/53 versus 12/51). In a subgroup of 15 patients who failed IVF with insufficient numbers of motile sperm, fertilization was significantly higher after subzonal sperm insertion. Partially zona-dissected embryos from couples with severe teratozoospermia (less than or equal to 5% normal forms; strict criteria) had significantly more morphological abnormalities than those from patients with moderate teratozoospermia (6% to 10% normal forms). In severely teratozoospermic patients, significantly fewer partially zona-dissected than subzonally inserted embryos implanted. CONCLUSIONS: The decision of which micromanipulation method to perform can possibly be based on careful analysis of sperm morphology.
Embryo Implantation↗
Relationship of follicular fluid prorenin to oocyte maturation, steroid levels, and outcome of in vitro fertilization.
Prorenin (PR) is present in high concentrations in the follicular fluid (FF) of the preovulatory follicle. It is the predominant form of renin detected in FF. Its biosynthesis and secretion from the ovary are regulated by gonadotropins. In the present study we measured PR and steroid levels in FF from 136 follicles. Follicular fluids were obtained, 36 h after hCG injection, from 41 ovarian-stimulated patients who underwent follicle puncture and oocyte retrieval for in vitro fertilization. We related FF PR to steroid levels and to the stage of oocyte-cumulus complex maturation. PR levels in 62 FF containing mature healthy fertilized oocytes averaged 2620 +/- 157 (+/- SE) ng/mL.h (728 +/- 44 ng/L.s; range, 1020-6880 ng/mL.h, 283-1911 ng/L.s). A subgroup of 16 of these follicles containing mature oocytes were from 7 women who conceived, in which PR levels spanned only the lower range from 1030-2720 ng/mL.h (286-756 ng/L.s). No patient conceived with FF PR above 2800 ng/mL.h (778 ng/L.s), yet one third of all mature follicles were above this range. Lower levels of PR were detected in FF containing immature oocytes (germinal vesicle stage) associated with either compact (1665 +/- 480 ng/mL.h; 463 +/- 133 ng/L.s; n = 22; P less than 0.02) or expanded (1785 +/- 193 ng/mL.h; 496 +/- 54 ng/L.s; n = 24; P less than 0.005) cumulus mass; a subgroup (n = 5) of follicles with immature oocytes and compact cumulus had very high levels of FF PR, ranging from 3830-7520 ng/mL.h (1064-2089 ng/L.s), while the remainder had levels less than 1300 ng/mL.h (361 ng/L.s). Progesterone and estradiol (E2) were lower in FF surrounding immature oocytes associated with compact (P less than 0.005) or expanded (P less than 0.02) cumulus, than in those containing mature oocytes. Testosterone (T) and androstenedione were measured in only a fraction of the samples; there were no apparent differences between follicles containing mature and immature oocytes. However, T and androstenedione levels were high in the subgroup of follicles containing immature oocytes and very high levels of PR. Of the hormones measured, T revealed the most striking relationship with PR (r = 0.62; n = 49; P less than 0.001).(ABSTRACT TRUNCATED AT 400 WORDS)
Androstenedione↗
Radioimmunoassay of inhibin based on synthetic human inhibin alpha-chain peptide.
Polyclonal rabbit antisera were produced against cyclic human inhibin [(Cys6, Tyr7) alpha-(6-30)NH2] peptide, covalently conjugated to bovine serum albumin. The tyrosine residue introduced at position 7 facilitated the oxidative incorporation of radiolabel (125I) to yield a tracer with specific activity of 73.9 Ci/g. These reagents were used to develop a homologous equilibrium radioimmunoassay for human inhibin, with polyethylene glycol, 200 g/L, serving as the separation phase. At a detection limit of 2 micrograms/L (n = 7), immunoactive inhibin was detectable in human pre-ovulatory follicular fluid (128 micrograms/L), seminal plasma (2374 micrograms/L), amniotic fluid (66 micrograms/L), and placental extract (347 micrograms/L). We also demonstrated inhibin immunoreactivity in biological fluids from other mammalian species: macaque, chimpanzee, porcine, and bovine, but not rodent (guinea pig). Although the antisera were raised against a nonbioactive inhibin peptide, immunoglobulins fractionated on Protein A-Sepharose neutralized the bioactivity of human ovarian inhibin. Further characterization of inhibin immuno- and bioactivity was undertaken with immobilized heparin, divalent metal cations, and dye ligands. Only heparin-Sepharose distinguished between immuno- and bioactive inhibin.
Amniotic Fluid↗
Pregnancy achieved through in vitro fertilization with cryopreserved semen from a man with Hodgkin's lymphoma.
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Adult↗