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ABSTRACT: The bi-parental genetic state is not a given after assisted reproduction. This is based on a biological definition of parentage that concerns generational inheritance of genetic material. Often three or more individuals may participate in artificial reproduction. Only cytoplasmic and spindle transfer can result in the genetic tri-parental state. All other forms involving three or more assisting persons with no heritable genetic contribution must be considered differently. Can a cytoplasmic donor be a biological parent based on a potential contribution of mitochondrial DNA to the offspring? - only if the mitochondrial DNA sequence can be traced back to the donor, a phenomenon which may not be very common. When considering spindle transfer for avoiding transmission of mitochondrial disease, all offspring is likely to be tri-parental.
Reproductive biomedicine online 03/2013; · 2.04 Impact Factor
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ABSTRACT: The ultimate goal of IVF is to achieve healthy, single, live births following each single-embryo transfer. A timeline for this eventuality has never been defined. National implantation rates from 2003-2010 provided by the Society for Assisted Reproductive Technologies (SART) in the USA were evaluated. Regression analysis was applied to the annual trends. A high correlation was noted showing a linear increase from year to year ranging between 0.3% and 1.5% when maternal age was not higher than 42. This relationship can be retrospectively applied to earlier SART data reports. This incline may be partly technology driven and resembles Moore's law, which describes annual improvements in microchip performance. Based on the assumption that technology will continue to drive progress, the length of time required to reach 100% implantation was calculated. The interval varied between 43years (AD 2053) for the youngest age group (<35years old) and 294years for the 41-42-year age group. The timeframe is shifted for the younger patients to an earlier date of 2027 if a subset of clinics with high implantation regression slopes and low variance is selected. The implications of these findings for infertility treatment and fertility preservation are discussed. Success after IVF has steadily improved. Data from US-based clinics are annually collected by the Society for Assisted Reproductive Technologies (SART; www.sart.org). Through SART, individual clinic's outcomes may be assessed. Although live birth and pregnancy are considered the gold standard of success, the investigators took the approach that those outcomes are often biased due to transfer of multiple embryos. The present analysis was therefore performed on individual embryos, by using the implantation rate to compare national and individual clinic datasets. National implantation rates show a linear increase from year to year ranging between 0.3% and 1.5% for patients aged <43years. We postulate that this linear trend can be traced back to 1985 even though statistical analysis could only be applied to the implantation data from 2003-2010. We expect that this annual incline is partly technology driven. This is an intriguing effect also seen in the computer industry where there has been a doubling of computer speed and memory for the past 47years, a phenomenon anticipated by Moore's law. We predict that the annual increase in implantation will also continue as new technologies become available. Based on current trends, the length of time for 100% implantation rates was calculated. Time to achieving 100% implantation varied between 43years (AD 2053) for the youngest age group (<35years old) to 294years for women 41-42years old. Some clinics may report a perfect success earlier than others. However, implantation does not guarantee birth.
Reproductive biomedicine online 09/2012; · 2.04 Impact Factor
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ABSTRACT: To determine whether a mandatory single-blastocyst transfer (mSBT) algorithm reduced multiple gestation rates without sacrificing clinical pregnancy rates.
Retrospective review.
U.S. university-based assisted reproductive technology (ART) program.
All women younger than 38 years undergoing their first ART cycle from 2009 to 2010 with ≥4 high-grade embryos on day 3 after oocyte retrieval (patients from 2009 were the "before" group, and patients completing ART under the mSBT policy in 2010 were the "after" group).
mSBT algorithm.
Multiple gestation and clinical pregnancy rates.
Of the qualified patients, 136 women met inclusion criteria (62 from 2009, 74 from 2010). The baseline demographics were similar between the groups. Statistically significantly fewer blastocysts were transferred per patient in 2010 compared with 2009 (1.5 vs. 1.9). The clinical pregnancy rates before (67.7%) or after (63.5%) the mSBT policy were not statistically significantly different. Multiple gestation rates were statistically significantly reduced, from 43.8% (2009) to 14.6% (2010) after the mSBT policy was instituted. More patients from 2010 had ≥1 blastocyst cryopreserved compared with 2009 (52.9% vs. 30.6%).
A novel single-blastocyst transfer algorithm reduced multiple gestation rates and improved cryopreservation rates without compromising clinical pregnancy rates in good-prognosis patients.
Fertility and sterility 07/2011; 96(3):580-4. · 3.97 Impact Factor
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ABSTRACT: Preimplantation genetic diagnosis has been proposed as a method to improve assisted reproduction technology outcomes, but different techniques have produced conflicting results. The use of appropriate techniques may provide positive outcomes.
Fertility and sterility 05/2009; 94(2):408-30. · 3.97 Impact Factor
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ABSTRACT: To evaluate trends and outcomes from preimplantation genetic diagnosis (PGD) cycles.
Retrospective data review.
A reference laboratory specializing in the provision of PGD services.
One hundred sixty-two patients at risk of transmitting a serious monogenic disorder to their children.
In vitro fertilization and PGD.
Results of PGD cycles.
Two hundred twenty-four PGD cycles were referred by 59 different IVF centers. Forty-six different disorders were diagnosed, including several not previously diagnosed at the preimplantation stage. Cystic fibrosis was the most common reason for referral (73 cases). A diagnosis was obtained for 84.4% of tested embryos, with results available 6 to 36 hours from sample receipt. Only 10.7% of cycles had no transfer. The pregnancy rate per cycle with ET was 43.4%.
Unlike previous reports of multiple PGD cycles, all of the cases in this study involved shipping of biopsied cells to a specialist reference laboratory for diagnosis. This approach, sometimes referred to as "transport PGD," accounts for the vast majority of PGD cycles in the United States. Preimplantation genetic diagnosis was shown to be an effective alternative to prenatal diagnosis for patients with an ethical or a religious objection to pregnancy termination and for infertile patients carrying a genetic disorder. Demand for this service at our center doubled in each of the last 4 years. Pregnancy rates per ET were encouraging, almost half of all patients undergoing their first PGD cycle achieving a birth or ongoing pregnancy.
Fertility and sterility 11/2008; 92(5):1544-56. · 3.97 Impact Factor
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ABSTRACT: Non-invasive methods of assessing embryo quality are critical for pregnancy success following IVF or intracytoplasmic sperm injection (ICSI). The addition of new non-invasive morphological and biochemical analyses may further improve pregnancy success, allowing the transfer of a single embryo, thereby reducing the risks involved in multiple births following IVF/ICSI. The presence of a protein, soluble human leukocyte antigen-G (sHLA-G), in embryo cultures has been suggested as a way to non-invasively predict embryo quality and pregnancy success, especially when used in conjunction with current embryo quality assessment methods. Detection of sHLA-G in embryo culture medium has been correlated with pregnancy success in 12 studies, but three studies were not able to detect sHLA-G. This is a review of the literature on sHLA-G detection in IVF/ICSI, and reasons are proposed for the reported discrepancies, as well as guidelines for reporting of data in future studies. Furthermore, it is suggested that the use of an HLA-G transgenic mouse model would advance understanding of the mechanism of action of sHLA-G in preimplantation embryos and its correlation to embryo health and viability. Research on a mouse model, combined with clinical studies, should enable the development of a fast and reliable method for utilizing sHLA-G detection to improve pregnancy success after IVF/ICSI.
Reproductive biomedicine online 11/2008; 17(4):470-85. · 2.04 Impact Factor
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ABSTRACT: This study examined a new method for vitrification of blastocysts that is safe, simple and easy to learn and use. Current vitrification techniques have shortcomings that include the use of dimethyl sulphoxide, one of the more toxic cryoprotectants, and minute containers that are difficult to handle and are usually open to contamination. Cell handling and loading times are very short, which allows no room for user-associated errors and increases the difficulty of the procedure. This study describes a method of vitrification without these shortcomings. Human and bovine blastocysts were exposed to a series of three cryoprotectant solutions and loaded into a 0.25 ml sterile straw, heat sealed at both ends and vitrified. This technique allowed sufficient time for cryoprotectant exposure, loading, sealing and vitrification. Research blastocysts were thawed, cultured for 24 h, and stained for cell viability. The majority survived and on average had few lysed cells. In clinical studies from three different centres, 81.4% of vitrified blastocysts were intact after thawing. Out of 43 transfers with 76 blastocysts replaced, 44.7% implanted, 43.4% yielded a fetal heart beat, and a total of 32 babies have been delivered or are ongoing. The overall clinical pregnancy per transfer rate was 60.4%. The high survival rates and clinical pregnancy rates obtained with this new, safe and easy-to-use vitrification procedure are encouraging.
Reproductive biomedicine online 10/2008; 17(3):360-7. · 2.04 Impact Factor
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ABSTRACT: To evaluate effects of luteinizing hormone (LH)-containing gonadotropins (human menopausal gonadotropin, hMG) on ploidy of human cleavage-stage embryos in gonadotropin releasing hormone (GnRH) antagonist cycles.
Retrospective matched cohort study.
Two academically affiliated private fertility centers, private preimplantation genetics laboratory, and medical school.
One hundred four consecutive in vitro fertilization cycles (IVF) with preimplantation genetic diagnosis in women aged 30 to 45 years.
Antagonist cycles with ovarian stimulation by either recombinant follicle stimulating hormone (FSH) alone, or in combination with human menopausal gonadotropin (FSH/hMG).
After matching patients for age and gonadotropin dosage, embryo ploidy, pregnancy, and miscarriage rates were evaluated.
Euploidy rates (FSH, 29.4% vs. FSH/hMG, 25.7%) and number of euploid embryos (FSH, 2.1 +/- 1.6 vs. FSH/hMG, 1.9 +/- 1.5) were similar between both groups, although trended in favor of FSH-only stimulation. FSH-only stimulation, however, demonstrated significantly higher clinical pregnancy rates per cycle start (FSH: 34.6% vs. FSH/hMG: 11.5%) and per embryo transfer (FSH: 40% vs. FSH/hMG: 15%).
Because this study involved mostly women of advanced reproductive age, at least in such an age category, LH-containing stimulation causes adverse effects on pregnancy rates, although whether such an adverse effect on ploidy exists in parallel requires further investigation. These observations point toward embryo-independent adverse effects on implantation (luteal phase) in antagonist cycles with hMG costimulation.
Fertility and sterility 10/2008; 92(3):937-42. · 3.97 Impact Factor
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ABSTRACT: The aim of this study was to evaluate the effect of ovarian stimulation with LH-containing gonadotropins (human menopausal gonadotropin, hMG), on ploidy of human cleavage-stage-embryos.
A total of 104 women, at ages 27-43 years, undergoing one cycle of controlled ovarian hyperstimulation for IVF in combination with preimplantation genetic diagnosis, were eligible for enrollment in this retrospective, controlled cohort study. Ovarian stimulation included down-regulation with long agonist and stimulation with either recombinant FSH or hMG. Since the ploidy of embryos changes with female age, patients were matched for age and dosage of the respective gonadotropin.
Despite similar numbers of chromosomally normal embryos in both groups, women undergoing hMG stimulation demonstrated significantly higher percentages of diploid embryos than did the FSH-stimulated patients (69.8 versus 45.3%; P < 0.01).
Long protocol LH-containing ovarian stimulation improves embryonic ploidy in comparison to pure FSH stimulation. This observation may explain higher IVF pregnancy rates, reported for hMG stimulation in some studies.
Human Reproduction 03/2008; 23(3):499-503. · 4.47 Impact Factor
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ABSTRACT: There is a known correlation between morphology and euploidy, but we believe that the association between the two embryo selection systems is too marginal to claim that it is absolute and that one can simply replace one system by the other. The optimal way to select euploid embryos is through a combination of morphology determination and preimplantation genetic diagnosis.
Fertility and sterility 11/2007; 91(3):943-5. · 3.97 Impact Factor
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New England Journal of Medicine 11/2007; 357(17):1769-70; author reply 1770-1. · 53.30 Impact Factor
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ABSTRACT: To improve preimplantation genetic diagnosis (PGD) accuracy by using "no result rescue" (NRR) consisting of the reanalysis of dubious results with additional probes binding to a locus different from the one previously analyzed.
Prospective study of PGD cycles with and without reanalysis of inconclusive results.
PGD laboratory.
Patients undergoing PGD for infertility or Robertsonian translocations.
Nuclei from day 3 biopsied embryos were analyzed with fluorescence in situ hybridization for chromosomes X,Y, 13, 15, 16, 17, 18, 21, and 22. When inconclusive results were obtained, NRR was performed. In addition, 100 PGD cycles using NRR were matched to controls according to maternal age, previous failed cycles, number of zygotes, number of eggs, and date of retrieval.
Determination of frequency of inconclusive results and error rate after use of additional probes. Comparison of frequency of inconclusive results with prior PGD results when NRR was not used. Assisted reproductive technology outcome was compared between PGD using NRR and controls not using PGD.
After analysis of 34,831 blastomeres from 34,225 embryos, 2,609 blastomeres (7.5%) showed inconclusive results. After NRR on those 2,609 blastomeres, the number of cells with inconclusive results was reduced to 3.1% (P<.001). After the introduction of NRR, fluorescence in situ hybridization errors, measured as discrepancies between the PGD diagnosis and the analysis of the nonreplaced embryo, decreased from 13.6% to 4.7% (P<.001). PGD with NRR significantly improved implantation rates, from 20% to 31%, and reduced spontaneous abortions from 27% to 6%.
The use of NRR has been proven to be a powerful tool to reduce the error rate and the frequency of inconclusive results in PGD, both parameters of high importance to assess quality of PGD laboratories. Indeed, these parameters are two of the few measurable criteria to measure PGD laboratories. In a parallel controlled study, PGD with NRR significantly improved implantation rates and reduced spontaneous abortions, showing that PGD is more efficient in selecting embryos that will reach term.
Fertility and sterility 08/2007; 88(1):53-61. · 3.97 Impact Factor
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ABSTRACT: The age-related decline in female fertility has been attributed to a variety of causes including progressive oocyte depletion, meiotic irregularities and mitochondrial dysfunction. However, additional factors could potentially be involved. To explore this possibility, comprehensive analysis of gene expression in human oocytes, discarded following IVF procedures and segregated by age, was undertaken using microarray methods. These findings indicate that the expression of oocyte genes, in a variety of major functional categories including cell cycle regulation, cytoskeletal structure, energy pathways, transcription control, and stress responses, are influenced by maternal age. These results are corroborated by a complementary extensive study using mouse oocytes.
Reproductive biomedicine online 07/2007; 14(6):700-8. · 2.04 Impact Factor
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ABSTRACT: To evaluate whether differences in results between studies that involve preimplantation genetic diagnosis for chromosome testing are affected by technology, such as the number of cells to be biopsied or by differences in study design.
Evaluation of studies of aneuploidy testing according to the use of probes, fixation technology, error determination, and number of cells per embryo analyzed.
Preimplantation genetic diagnosis laboratories.
Patients in published studies who underwent preimplantation genetic diagnosis for infertility or repeated pregnancy loss.
As determined by each evaluated study, the number of biopsied cells and its effect on further development was evaluated by a comparison of models of embryo freezing and partial cell loss.
Use of probes, fixation strategy, number of biopsied cells, and error rate determination of different published studies.
Differences in results between studies can be explained by the technology used and are not affected necessarily by differences in design and patient allocation.
Studies that contradict the finding that aneuploidy screening improves implantation and lowers miscarriage rates all have > or =1 of the following aspects in common: (I) an excess of cells having been removed; (II) inadequate choice of probes; and (III) suboptimal fixation technology.
Fertility and sterility 04/2007; 87(3):496-503. · 3.97 Impact Factor
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ABSTRACT: Human embryonic stem cells (hESC) have the potential to treat a wide range of diseases. Currently, the use of existing hESC lines in human clinical applications is limited, as they are derived from blastocysts subjected to immunosurgery with animal derived antibodies, and are maintained on mouse embryonic feeder (MEF) cells, in the presence of either fetal calf serum (FCS) or on Matrigel or with conditioned media from MEFs. Successful derivation of hESCs in xeno-free conditions is crucial in advancing stem cell therapy applications. Two hESC lines, one from chromosomally abnormal embryos and another cell line from normal embryos from the inner cell mass of human blastocysts are derived using a culture media that had 20% serum replacement (SR) and human FGF2 on human foreskin fibroblasts as feeder cells. Derivation and characterization of such xenofree hESCs suitable for clinical studies is described in this chapter.
Methods in molecular biology (Clifton, N.J.) 02/2007; 407:1-10.
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ABSTRACT: Intact zona-free human oocytes were screened using a combinatorial peptide library selection protocol. Pieczenik Peptide Sequence 1 (PPS1) HEHRKRG binds human spermatozoa. A complementary and unique binding sequence HNSSLSPLATPA (PPS2) was developed from the first PPS1 ligand that binds to the human zona pellucida or oolemma. Cytoplasm-free zonae from unfertilized eggs were obtained and used as an assay system to test the effects of exposure to these two ligands. Spermatozoa were inserted into evacuated zonae and their behaviour and binding activity were assessed at regular intervals. The behaviour of spermatozoa exposed to PPS1 and unlabelled spermatozoa injected into unexposed zonae was similar as far as binding was concerned (50 and 54% binding), but PPS1 exposed spermatozoa had higher motility and displacement, marked by their escape from the zona pellucida. Zonae exposed to PPS2 inhibited the interaction between injected spermatozoa and the inside of the zona when compared with controls (8.3 and 53.8% attached respectively, P < 0.001). The sperm-zona pellucida interaction described in this paper is applied as a functional assay for molecular interactions of sperm binding and can be used to assess function for potential surface markers on gametes. It is shown here that a unique binding ligand (PPS2) can be synthesized from another complimentary ligand (PPS1) without the need for a known intermediate substrate. PPS1 and PPS2 may have properties that can be used to target processes involved in conception and assisted reproduction. A movie sequence taken approximately 30 min after injection of spermatozoa into empty human zonae pellucidae shows behaviour of non-manipulated spermatozoa into zonae not exposed or exposed to ligand. This may be purchased for viewing on the Internet at www.rbmonline.com/Article/2159 (free to web subscribers).
Reproductive biomedicine online 10/2006; 13(3):361-7. · 2.04 Impact Factor
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ABSTRACT: Previous investigations revealed that choline-based freezing media developed in our laboratory were superior to conventional sodium-based media for storing mouse oocytes. This paper examines the ability of the choline-based medium CJ2 and a modified form of this medium, CJ3, to cryopreserve unfertilized human oocytes. Oocytes that were consented for research and matured overnight, as well as freshly collected, donor, mature metaphase II (MII) oocytes, were cryopreserved using choline-based media and an optimized slow-cooling protocol. The results showed higher survival and fertilization rates when CJ3 supplemented with 0.2 mmol/l sucrose was used as compared with CJ2 supplemented with either 0.1 mmol/l or 0.2 mmol/l sucrose. Freshly collected oocytes were more difficult to cryopreserve than those matured in vitro. Modification of the base medium proved to be one of the key factors in obtaining survival rates over 90%. Fertilization rates, embryo development, and genetic analysis of embryos resulting from control and frozen-thawed oocytes are provided. There appears to be a high correlation between chromosomal anomalies and abnormal morphology in embryos from thawed oocytes.
Reproductive biomedicine online 09/2006; 13(2):222-7. · 2.04 Impact Factor
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ABSTRACT: Several studies have shown a correlation between the pronuclear morphology score (PNMS) and subsequent embryo development and implantation. Embryos with poor pronuclear score, elsewhere referred to as Z3 and Z4, are often not transferred or cryopreserved because it is believed that they have poor pregnancy potential. The objective of this study is to report our data on the use of the pronuclear score and its effect on pregnancy outcome.
Retrospective analysis of IVF/ICSI-embryo transfer cycles completed over the course of 1 year (n = 334). Comparisons were made only in those groups of patients in whom cohorts of similarly scored PNMS embryos were transferred. The proportion of such homologous cohorts was 104/334 (31%). All other replacements were excluded from final analysis as they were dissimilar as far as PNMS is concerned. Pregnancy outcomes were evaluated.
The incidence of live birth resulting from the transfer of single pronuclear score homologous embryo types was 56 (14/25), 41 (13/32), 54 (23/43) and 0% (0/4) for PNMS scores 1, 2, 3 and 4, respectively. There was no correlation between PNMS category of the embryos transferred and live birth rates (P = 0.139).
PNMSs of 1, 2 or 3 do not correlate with live birth rates when assessing unique PNMS embryo transfers. In particular, previously considered poor (type 3) embryos can result in pregnancy with normal live birth rates. Whether type 4 embryos are compatible with normal development remains to be shown.
Human Reproduction 07/2006; 21(6):1599-604. · 4.47 Impact Factor
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Fertility and sterility 05/2006; 85(4):1071. · 3.97 Impact Factor
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ABSTRACT: The inicidence of miscarriage is correlated with maternal age. The majority of miscarriages are chromosomally abnormal. The purpose of this study was to determine in a large population of infertility patients (>2000 cycles) if preimplantation genetic diagnosis (PGD) reduced the rate of spontaneous abortions.
Multicenter retrospective controlled study.
One hundred IVF centers referring samples to a reference PGD laboratory.
Infertile women.
The spontaneous abortion rate after PGD was retrospectively compared to non-PGD cycles from the 2002 American Society for Reproductive Medicine-Society for Assisted Reproduction Technology report on IVF cycles.
Spontaneous abortions and trisomic offspring rates.
The study included 2,279 cycles of PGD. The pregnancy rate per retrieval was 26.7% (average age 39.6). The mean pregnancy loss for the PGD group (0.167) was significantly lower than for the general IVF group (0.215) (P<.001). After PGD, the spontaneous abortion rate was 14.1% for ages 35-40, and 22.2% for women over 40, compared to 19.4% (P=.03) and 40.6% (P<.001), respectively, in controls. The clinical error rate of PGD (1.2%) was significantly lower than expected (4.7%) (P<.001).
The data suggests that PGD significantly reduces the risk of spontaneous abortions in women undergoing IVF and PGD, particularly in women over 40. In addition, PGD may also reduce the risk of trisomic offspring.
Fertility and sterility 02/2006; 85(2):326-32. · 3.97 Impact Factor
