Background: The general trend in PGD is the analysis of higher numbers of chromosomes requiring up to 2 rehybridizations. However, there may be diminishing returns as the increase in the number of chromosomes analyzed will result in a minimal increase in sensitivity and a greater decrease in specificity (may yield a higher number of indeterminate or false-positive abnormal embryos). Objective: The purpose of this study was to evaluate the percent of embryos found to be normal out of total number of embryos biopsied in IVF cycles employing PGD using a 5 chromosome panel vs. a 9 chromosome panel and the effect on pregnancy rates. Materials and Methods: This was a retrospective analysis of 226 patients undergoing IVF with PGD from September 2003 to May 2004. The medical records were reviewed for age, number of embryos biopsied, number of normal embryos per PGD, number of embryos transferred, and pregnancy outcome for 4 physicians with similar stimulation protocols. 55 patients had necessary information documented and were used for the study. Student t-test for two samples and chi-square test were used for statistical analyses. Results: The data for the two groups is shown in Table 1. The mean age, number of total embryos biopsied and number of embryos transferred were similar between the two groups. The 5 chromosome groups had higher number and percent of normal embryos as diagnosed by PGD (p<0.005, p<0.0001). The pregnancy rate was higher in the 5 chromosome group, but this did not reach significance. The patients able to conceive (5 or 9 chromosome panel) had a significantly higher number of normal embryos as diagnosed by PGD (4.0 ± 2.2 vs. 2.0 ± 0.97, p<0.0005) and a higher number of embryos transferred (2.5 ± 0.7 vs. 1.9 ± 0.7, p<0.005) as compared to those unable to conceive. Tabled 1Means5 Chr PGD9/10 Chr PGDAge3637# Embryos biopsied7.366.73# Normal embryos4.02.4%Normal embryos6138# Embryos transferred2.322.15Pregnancy rate68%50% Open table in a new tab Conclusions: A trend towards higher pregnancy rates was noted in the 5 chromosome PGD group. This may indicate that with increased number of chromosomes analyzed there is a greater decrease in specificity with a higher number of false-positive abnormal embryos, resulting in fewer embryos available for transfer. Therefore, PGD using a 9 chromosome panel offers little added value over the 5 chromosome panel with a potential adverse effect on pregnancy rates.

The aim of this study was to determine if day 3 embryos derived from eggs of unknown fertilization status (no observable pronuclei) determined at 16-18 hours post-insemination or ICSI had increased rates of aneuploidy over those having observable pronuclear formation within the normal “window” for such observation. This was a retrospective analysis of data collected from 226 patients who requested preimplantation genetic diagnosis-aneuploidy screening (PGD-AS). The charts were reviewed for morphological grading of both zygotes and embryos as well as their ploidy. Fifty-four patients with a total of 76 embryos met inclusion criteria. Out of the 76 embryos that developed from eggs with no observable pronuclei, 20 embryos were either not biopsied due to poor development or the PGD-AS results were inconclusive. Seventeen (30.4%) of the remaining embryos were normal, while 39 (69.6%) were abnormal as diagnosed by PGD. Seven of the normal embryos developed from 1PN “zygotes”, one developed from a 3PN zygote and 9 developed from either meiosis I or meiosis II oocytes (Table1, Figure 1). Table 1 Tabled 1 This data suggests that embryos developing from cells in which a 2PN status is never identified, are often poor quality and are mostly abnormal as identified by PGD. Those normal embryos that developed from 1PN “zygotes” might have been read to early or too late and the 2PN stage might have been missed, suggesting that more frequent reading of the embryos should be considered.

Background: ICSI (Intracytoplasmic Sperm Injection) is usually performed in couples with male factor infertility on the day of the oocyte retrieval. PGD (Preimplantation Genetic Diagnosis) is performed for single gene disorders or aneuploidy testing. Objective: To describe a case in which embryos were treated by rescue ICSI in combination with PGD from which a pregnancy resulted. Materials and Methods: A case report is described. The patient is a 43 year old Gravida 2 Para 0020 with infertility with her present partner of one and half year duration. Her past OB/GYN history is significant for a spontaneous abortion with a biochemical pregnancy in 1988, as well as a therapeutic abortion in 1987. Her husband is a healthy 35 years old with previous proven fertility with a different partner several years ago. His semen analysis was obtained 8 months prior to the current IVF cycle and was deemed normal. The patient was stimulated in the following manner. Oral contraceptives were administered, followed by ovarian stimulation with gonadotropins along with a GnRH Antagonist. HCG was administered when follicle maturity was determined and 12 oocytes retrieved 36 hours later. On the day of retrieval, specimen collection from the patient's husband resulted in 2.4ml of semen with 70 million/ml and 50% sperm motility rate. Oocytes were inseminated 4 h after retrieval. Results: On the following day no fertilization was observed and rescue ICSI was therefore performed on the 7 mature eggs, of which 2 normally fertilized. The patient desired PGD and therefore single blastomeres were then biopsied from 2 embryos 4 days after retrieval. FISH analysis was performed for chromosomes 13, 18, 21, X and Y. One embryo was diagnosed as normal and the other with trisomy 21. On day 5 the normal embryo was a 10 cell stage and was transferred into the patient's uterus. The patient was continued on Progesterone supplementation and 2 weeks later the patient's pregnancy test was positive with an hCG level of 560 mIu/ml. An intrauterine singleton pregnancy was then confirmed several weeks later by sonography. Conclusion: Rescue ICSI in combination with PGD can result in successful pregnancy.

The aim of the study was to determine if embryo morphology on day 3 correlates with aneuploidy diagnosis by PGD (preimplantation genetic diagnosis). Retrospective A retrospective analysis was performed on data collected from consecutive patients requesting aneuploidy screening of their embryos over a twelve month period. Morphological grading of embryos as well as their PGD results was recorded. Out of 1538 embryos biopsied from two hundred and twenty six patients, 326 (21%) were <6 cells and 1212 (79%) were >6 cells at the time of biopsy on day three. Two hundred and twelve (or 65%) of these embryos were aneuploid vs. 493 (or 41%) in those ≥ 6 cells (table 1), a statistically significant difference (p<.001). Further analysis of these embryos revealed that in the embryos less than 6 cells, with grades A or B, 60/150 or 40% were normal versus 28/150 or 19% if the embryos had grades C or D (fig 1) and the difference was statistically significant (p<.001). Tabled 1 View Large Image Figure ViewerDownload (PPT) This data suggests that morphology can have a predictive value of aneuploidy status and that slow-cleaving, morphologically compromised embryos are almost always aneuploid. As these embryos invariably arrest upon extended culture, not performing PGD on this subset of embryos poses little risk of discarding healthy, euploid embryos capable of establishing a sustained pregnancy, and has the added benefit of conserving valuable medical resources.

Background: PGD (preimplantation genetic diagnosis) for the purpose of aneuploidy screening (AS) has increased in usage over the past decade and is usually performed 3 days after oocyte retrieval. Our ART program currently performs AS on approximately twenty percent of patients requiring IVF-ET, and embryo biopsy was typically performed on virtually every embryo generated from these patients, regardless of their rate of cleavage or "grade" at biopsy. Objective: The aim of the study was to determine if slow cleaving, poor grade embryos are more likely to be chromosomally abnormal, and if biopsy of these embryos is either of value or is in fact a waste of medical resources. Materials and Methods: A retrospective analysis was performed on data collected from consecutive patients requesting aneuploidy screening of their embryos over a five month period. Morphological grading of embryos as well as their PGD results was recorded. Results: Out of 683 embryos biopsied from one hundred and eleven patients, 113 (16%) were < 6 cells at the time of biopsy on day three. Seventy four of 113 (or 65%) of these embryos were aneuploid. Further analysis of these embryos revealed a roughly equal distribution of normal versus aneuploid (26 vs. 30) outcomes if the grade of the embryos was in the A-B (good) range. In contrast, slow-cleaving embryos in the C-D (poor) morphologic range were five times as likely (11 normal vs. 55 aneuploid) to be abnormal in their chromosomal makeup, as well as three times as likely to have anucleated blastomeres (16 of 22) as did good grade, albeit slow-cleaving embryos. Conclusion: This data suggests it is reasonable to biopsy <6 cell embryos on day three post-insemination if they are of good morphology. Slow-cleaving, morphologically compromised embryos are almost always aneuploid, and as these embryos invariably arrest upon extended culture, not performing AS on this subset of embryos poses little risk of discarding healthy, euploid embryos capable of establishing a sustained pregnancy, and has the added benefit of conserving valuable medical resources.

Background: Preimplantation genetic diagnosis (PGD) is often performed in patients with genetic disorders to prevent transmission to offspring. Spinocerebellar Ataxia Type 3 (SCA3) is a rare autosomal dominant disease. It is characterized by progressive cerebellar ataxia and variable findings including a dystonic-rigid syndrome, a Parkinsonian syndrome or a combined syndrome of dystonia and peripheral neuropathy. The diagnosis of SCA3 rests upon the use of DNA-based testing to detect an abnormal CAG trinucleotide repeat expansion of the Machado-Joseph disease (MJD) gene. Objective: To describe a case report in which PGD in was utilized in a woman with Spinocerebellar Ataxia Type 3 in order to decrease offspring transmission of the disease. Materials and Methods: This is a case report of a woman affected with SCA3. The patient's mother had developed SCA3, so DNA-based testing was performed on the patient revealing 73 CAG repeats on one of the alleles of the MJD gene confirming the diagnosis of SCA3. In an effort to avoid passing the rare gene to her offsprings, PGD analysis was performed specifically selecting for SCA3 on the MJD gene. Results: The patient underwent two IVF cycles. In the first cycle, 11 oocytes were aspirated, and ICSI (Intracytoplasmic Sperm Injection) was performed on the 6 that appeared mature. On PGD analysis, 2 embryos yielded inconclusive results, 3 embryos were affected with SCA3, and 1 embryo was not affected however displayed monosomy 22. Therefore, no normal embryos were identified on the first cycle and no embryo transfer took place. In the second cycle, 18 oocytes were aspirated, 10 of which appeared mature for ICSI. Of these, 5 were affected, 3 were unable to be studied and 2 were normal after PGD analysis. These 2 embryos were subsequently transferred on day 6. Pregnancy was confirmed 10 days after the transfer and currently the patient carries a normal twin pregnancy. Conclusions: A specific gene was selected for with PGD analysis in IVF and embryos not carrying the selected mutation were selected resulting in normal twin pregnancy. PGD can be utilized to select for specific gene mutations known to be present in the family.

Objective: The purpose of this Study was to determine the positive predictive value (PPV) and negative predictive value (NPV) of FISH analysis and to determine which chromosomal abnormalities are most frequently confirmed.Design: Prospective observational.Setting: IVF laboratory.Patient(s): Two hundred forty-one embryos were analyzed from 98 patients.Intervention(s): FISH reanalysis.Main Outcome Measure(s): Embryos that would have been discarded in patients undergoing preimplantation genetic diagnosis (PGD) were fixed and FISH reanalysis was performed. Results of reanalysis were compared with the day 3 diagnosis while PPV and NPV were calculated.Result(s): Among the 241 embryos, 198 embryos were abnormal and 43 were normal by day 3 FISH analysis. The PPV was 83% and the NPV was 81%. PPV was also determined for specific categories of aneuploidy, and certain abnormalities such as monosomies, trisomies, tetrasomies, and polyploidies were frequently confirmed on reanalysis (PPV >80%), whereas Turner syndrome diagnosis was not (PPV = 17%).Conclusion(s): FISH analysis offers a PPV of 83% and NPV of 81% when evaluating a single blastomere in conjunction with PGD. FISH errors and mosaicism are primarily responsible for the errors associated with FISH analysis in PGD.

ObjectiveThe purpose of this study is to determine the chromosomal make-up of those embryos diagnosed as abnormal by PGD (Preimplantation Genetic Diagnosis).DesignProspective.Materials and methodsAll embryos determined to be abnormal by PGD, that would have otherwise been discarded, were included in this study. These abnormal embryos were fixed one or two days after PGD and FISH analysis was performed on all nuclei that were fixed. Fifty-six embryos were included in the study from 27 IVF/ICSI patients that had PGD. Single blastomeres were biopsied on day 3, 4–12 cell stage embryos. Chromosomes 13, 18, 21, X and Y were analyzed by Fluorescence in Situ Hybridization (FISH). Euploid embryos (on average 2.5/patient) were transferred back on day 4–6 post-retrieval or cryopreserved. The abnormal embryos, that would have otherwise been discarded, were used for this study after patient consent was obtained.ResultsOut of the 56 abnormal embryos, on FISH confirmation, the day 3 diagnosis was confirmed in 29 and 10 embryos were deemed as normal on reconfirmation (the criteria of normal: if the embryo has more than 50% normal cells) (graph 1). However out of these embryos, on reanalysis, 3 had all normal nuclei, while the remaining ones were mosaic. The remaining 17 embryos had a different abnormality or a combination of abnormalities (e.g. a monosomic embryo on day 3 may have trisomic result on day 4). Of the embryos that had false positive results (diagnosed as abnormal by PGD when they were in fact normal), the most common abnormalities encountered were monosomies and trisomies and they were also the most frequent diagnoses that were confirmed Graph 2 and table 1). 6 chromosomally normal embryos were also analyzed for this study. These embryos would have otherwise been discarded and all 6 were confirmed as normal when FISH analysis was done day 4 or 5. Table 1ConclusionView Large Image Figure ViewerDownload Hi-res image Download (PPT) ObjectiveThe purpose of this study is to determine the chromosomal make-up of those embryos diagnosed as abnormal by PGD (Preimplantation Genetic Diagnosis). The purpose of this study is to determine the chromosomal make-up of those embryos diagnosed as abnormal by PGD (Preimplantation Genetic Diagnosis). DesignProspective. Prospective. Materials and methodsAll embryos determined to be abnormal by PGD, that would have otherwise been discarded, were included in this study. These abnormal embryos were fixed one or two days after PGD and FISH analysis was performed on all nuclei that were fixed. Fifty-six embryos were included in the study from 27 IVF/ICSI patients that had PGD. Single blastomeres were biopsied on day 3, 4–12 cell stage embryos. Chromosomes 13, 18, 21, X and Y were analyzed by Fluorescence in Situ Hybridization (FISH). Euploid embryos (on average 2.5/patient) were transferred back on day 4–6 post-retrieval or cryopreserved. The abnormal embryos, that would have otherwise been discarded, were used for this study after patient consent was obtained. All embryos determined to be abnormal by PGD, that would have otherwise been discarded, were included in this study. These abnormal embryos were fixed one or two days after PGD and FISH analysis was performed on all nuclei that were fixed. Fifty-six embryos were included in the study from 27 IVF/ICSI patients that had PGD. Single blastomeres were biopsied on day 3, 4–12 cell stage embryos. Chromosomes 13, 18, 21, X and Y were analyzed by Fluorescence in Situ Hybridization (FISH). Euploid embryos (on average 2.5/patient) were transferred back on day 4–6 post-retrieval or cryopreserved. The abnormal embryos, that would have otherwise been discarded, were used for this study after patient consent was obtained. ResultsOut of the 56 abnormal embryos, on FISH confirmation, the day 3 diagnosis was confirmed in 29 and 10 embryos were deemed as normal on reconfirmation (the criteria of normal: if the embryo has more than 50% normal cells) (graph 1). However out of these embryos, on reanalysis, 3 had all normal nuclei, while the remaining ones were mosaic. The remaining 17 embryos had a different abnormality or a combination of abnormalities (e.g. a monosomic embryo on day 3 may have trisomic result on day 4). Of the embryos that had false positive results (diagnosed as abnormal by PGD when they were in fact normal), the most common abnormalities encountered were monosomies and trisomies and they were also the most frequent diagnoses that were confirmed Graph 2 and table 1). 6 chromosomally normal embryos were also analyzed for this study. These embryos would have otherwise been discarded and all 6 were confirmed as normal when FISH analysis was done day 4 or 5. Table 1 Out of the 56 abnormal embryos, on FISH confirmation, the day 3 diagnosis was confirmed in 29 and 10 embryos were deemed as normal on reconfirmation (the criteria of normal: if the embryo has more than 50% normal cells) (graph 1). However out of these embryos, on reanalysis, 3 had all normal nuclei, while the remaining ones were mosaic. The remaining 17 embryos had a different abnormality or a combination of abnormalities (e.g. a monosomic embryo on day 3 may have trisomic result on day 4). Of the embryos that had false positive results (diagnosed as abnormal by PGD when they were in fact normal), the most common abnormalities encountered were monosomies and trisomies and they were also the most frequent diagnoses that were confirmed Graph 2 and table 1). 6 chromosomally normal embryos were also analyzed for this study. These embryos would have otherwise been discarded and all 6 were confirmed as normal when FISH analysis was done day 4 or 5. Table 1 Conclusion

Zona drilling by Computer Assisted Infrared Laser is easy and precise. However the safety of laser used in IVF laboratory is still a concern. This study aims to evaluate the embryo biopsy procedure and to compare two different biopsy techniques: the Computer Assisted Infrared Laser (CAIL), using the Hamilton Thorn-Zilos-tk laser and the Acid Tyrode's Solution (ATS). Retrospective A total of 233 patients that underwent PGD for aneuploidy screening between 04/01/03 to 3/15/2004 were included in the study. The laboratory environment including the culture system, lab personnel and clinical protocols were identical in all cases. All patients had ovulation induction with gonadotropins and HCG was administered when follicle maturity was determined. Oocyte retrieval was performed 36 hours after HCG and single blastomeres were biopsied 3 days after retrieval. FISH analysis was performed for chromosomes 13, 18, 21, X and Y. 137 PGD cases (average age 38.72) had embryo biopsy preceded by ATS while 96 cases (average age 38.93) had CAIL zona opening before embryo biopsy. In addition 616 IVF patients (average age 38.56) that did not have PGD were included as controls. In both groups, blastomeres were rarely damaged. In the ATS group 123/144 cases had embryo transfers and the average number of embryos transferred was 2.59. In the CAIL group, 78/96 cases had embryo transfers and the average number of embryos transferred was 2.22. The pregnancy rate between the ATS and CAIL group was not statistically different (45.5 and 49 % respectively). The embryo implantation was 40% in the ATS compared to 42% in the CAIL group with no significant difference. In addition when compared to the control group, there was no statistically significant difference in the pregnancy rate (40 and 42 % in the ATS and CAIL group and 38.5% in the control group) Computer assisted laser system is easier and leads to more precise zona opening. Blastomeres were rarely damaged in either group. The laser zona opening technique does not influence embryo development and results in similar pregnancy and implantation rates as the chemical method that uses Acid Tyrode's solution.

ObjectiveThe purpose of this study is to evaluate the pregnancy rate of patients who undergo PGD (Preimplantation Genetic Diagnosis) and to compare it to patients who have IVF (In Vitro Fertilization) without PGD.DesignRetrospective.Materials and methods912 patients that had IVF with or without PGD from 1/4/03 to 3/31/04 at our clinic were included in the study. Of these, 296 patients had PGD and 616 women had IVF without PGD. These patients had ovarian stimulation with gonadotropins and most received either GnRH agonists or antagonists as part of their stimulation protocol. Ovulation was induced with HCG when at least two follicles were mature, and oocyte retrieval was performed 36 hours after HCG. In the women that desired PGD, PGD was performed 3 days after retrieval and embryo transfer was performed day 3–6. Pregnancy rates were then determined by HCG level and were compared among the 2 groups. Statistical analysis was performed using the Fisher’s exact test.ResultsThe patient characteristics are seen in table 1. The average age for the 296 PGD patients was 38.76 and the average age for the 616 non PGD patients was 38.56. Of these, 52 patients in the PGD group had no embryos transferred, usually secondary to the fact that all embryos were diagnosed as abnormal. Among the IVF patients, 110 had no embryo transferred due to multiple reasons such as desire for cryopreservation, poor embryos or risk for ovarian hyperstimulation syndrome. When comparing the pregnancy rate in the patients that had embryo transfers, the pregnancy rate among the PGD and non-PGD patients did not vary significantly (45.08% vs. 38.54% respectively). Table 1 Patient CharacteristicsTable 2 Pregnancy rates in Embryo Transfer GroupsConclusionView Large Image Figure ViewerDownload Hi-res image Download (PPT) ObjectiveThe purpose of this study is to evaluate the pregnancy rate of patients who undergo PGD (Preimplantation Genetic Diagnosis) and to compare it to patients who have IVF (In Vitro Fertilization) without PGD. The purpose of this study is to evaluate the pregnancy rate of patients who undergo PGD (Preimplantation Genetic Diagnosis) and to compare it to patients who have IVF (In Vitro Fertilization) without PGD. DesignRetrospective. Retrospective. Materials and methods912 patients that had IVF with or without PGD from 1/4/03 to 3/31/04 at our clinic were included in the study. Of these, 296 patients had PGD and 616 women had IVF without PGD. These patients had ovarian stimulation with gonadotropins and most received either GnRH agonists or antagonists as part of their stimulation protocol. Ovulation was induced with HCG when at least two follicles were mature, and oocyte retrieval was performed 36 hours after HCG. In the women that desired PGD, PGD was performed 3 days after retrieval and embryo transfer was performed day 3–6. Pregnancy rates were then determined by HCG level and were compared among the 2 groups. Statistical analysis was performed using the Fisher’s exact test. 912 patients that had IVF with or without PGD from 1/4/03 to 3/31/04 at our clinic were included in the study. Of these, 296 patients had PGD and 616 women had IVF without PGD. These patients had ovarian stimulation with gonadotropins and most received either GnRH agonists or antagonists as part of their stimulation protocol. Ovulation was induced with HCG when at least two follicles were mature, and oocyte retrieval was performed 36 hours after HCG. In the women that desired PGD, PGD was performed 3 days after retrieval and embryo transfer was performed day 3–6. Pregnancy rates were then determined by HCG level and were compared among the 2 groups. Statistical analysis was performed using the Fisher’s exact test. ResultsThe patient characteristics are seen in table 1. The average age for the 296 PGD patients was 38.76 and the average age for the 616 non PGD patients was 38.56. Of these, 52 patients in the PGD group had no embryos transferred, usually secondary to the fact that all embryos were diagnosed as abnormal. Among the IVF patients, 110 had no embryo transferred due to multiple reasons such as desire for cryopreservation, poor embryos or risk for ovarian hyperstimulation syndrome. When comparing the pregnancy rate in the patients that had embryo transfers, the pregnancy rate among the PGD and non-PGD patients did not vary significantly (45.08% vs. 38.54% respectively). Table 1 Patient CharacteristicsTable 2 Pregnancy rates in Embryo Transfer Groups The patient characteristics are seen in table 1. The average age for the 296 PGD patients was 38.76 and the average age for the 616 non PGD patients was 38.56. Of these, 52 patients in the PGD group had no embryos transferred, usually secondary to the fact that all embryos were diagnosed as abnormal. Among the IVF patients, 110 had no embryo transferred due to multiple reasons such as desire for cryopreservation, poor embryos or risk for ovarian hyperstimulation syndrome. When comparing the pregnancy rate in the patients that had embryo transfers, the pregnancy rate among the PGD and non-PGD patients did not vary significantly (45.08% vs. 38.54% respectively). Table 1 Patient Characteristics Table 2 Pregnancy rates in Embryo Transfer Groups Conclusion

Six hundred sixty human embryos from 94 IVF/ICSI cycles were biopsied on Day 3 and a single blastomere removed for identification of the five chromosomes 13, 18, 21, X and Y (those most involved in human aneuploidy) by Fluorescent in Situ Hybridization (FISH). 367(55.6%) of the embryos were euploid and 213 embryos were transferred in 85 patients. The average number of embryos transferred was 2.5 per patient. 44 patients were pregnant (51.8%/ET). 281 (42.6%) embryos were diagnosed as aneuploid. 86 normal embryos left over from transfer and all 281 aneuploid embryos were further cultured to Day 6. Aneuploid embryos are more likely to arrest (73.7%) before day 6 than normal embryos (37.2%) at extended culture (p<0.01). In the normal group, 54(62.8%) went to blastocyst/morula, whereas 74(26.3%) in the aneuploid group developed to blastocyst (56, 19.9%)/morula. Chromosomal reanalysis for the same probes of the Inner Cell Mass of the aneuploid group blastocysts confirmed that 34(60.7%) were aneuploid (trisomy, polyploidy or monosomy) that were concordant from D3 single cell diagnosis, 22(39.3%) of the blastocysts were diploid. The reason for this non-predictable outcome is due not only to FISH errors such as overlapped signals, hybridization failure, or nuclear material lost during either slide fixation or FISH. We believe mosaicism of the D3 embryo is a significant cause of this non-concordance and that the embryo has limited self-correction abilities in its development.

Introduction: Two couples with multiple miscarriages were referred for PGD following diagnosis of the reciprocal translocation in the male partners: 46,XY t (2,7)(p11.2;q22.1) and 46,XY t(2,7)(p23.1;p22.1). Objective: Using single interphase cell FISH with chromosome-specific breakpoint spanning probes PGD to assist translocation carriers to have babies. Materials and Methods: Before PGD, lymphocytes were used to test the efficiency of the probes. Sperm were analyzed by FISH using the same probe sets to predict the chromosome status of the embryos. Embryo biopsies were performed on Day 3 after fertilization. Single blastomeres were biopsied. Embryo biopsy of case 1 was done by ART personnel in Dr. Vermesh’s IVF lab. Embryo biopsy of case 2 was done by Dr. Yang in Florida and slides were shipped to ART for translocation diagnosis. Chromosome-specific breakpoint spanning probes were used in FISH. Three embryos were transferred in both cases. Result: Tabled 1Normal/balanced Sperm percentageTotal embryosNormal/balanced embryosEmbryos transferredPregnancyAmniocentesis resultCouple 147%733Triplets2 balanced 1 normalCouple 225%1843SingletonNot done yet Open table in a new tab Conclusion: PGD using single interphase blastomere FISH for translocation carriers by breakpoint spanning probes can help patients have phenotypically normal offspring.

With the improvement of in vitro culture system and optimized embryo development, implantation rate has been improved dramatically. Implantation rates as high as 70% have been reported when morphologically good blastocysts were transferred (DK Gardener, 2004). However, the number of advanced maternal age patients has increased significantly, and therefore have the number of trisomies. The aim of this study was to compare the pregnancy outcomes of blastocyst transfer cases in PGD vs. non-PGD cycles. Retrospective. A total of 386 patients that underwent blastocyst transfer were included in the study. All patients had ovulation induction with gonadotropins and HCG was administered when follicle maturity was determined. Oocyte retrieval was performed 36 hours after HCG and single blastomeres were biopsied 3 days after retrieval. FISH analysis was performed for chromosomes 13, 18, 21, X and Y. 349 blastocyst transfer cases without PGD aneuploid screening (Non-PGD group, average age 34.10) while 37 cases (PGD group, average age 36.49) had blastocyst transfer after PGD-AS (PGD-Aneuploidy Screening). In the Non-PGD group, pregnancy rate was 51.3% (179/349) with the average 2.59 embryos transferred. In the PGD group, pregnancy rate was higher 64.9% (24/37) with less embryos transferred (average number of embryos transferred was 2.22) in older patient (36.49 vs. 34.10), although the difference was not significant. The embryo implantation was 50% in the PGD group compared to 47.8% in the non-PGD group with no significant difference. Table 1. Pregnancy outcomes after blastocyst transfer in PGD vs. non-PGD groups: *compared to PGD-AS group, the two-tailed Fisher’s exact test p value equals 0.1230, no significant difference. With the improvement of in vitro embryo culture system and use of sequential culture media, more embryos develop to the blastocyst stage. PGD-AS combined with morphology selection can improve single blastocyst transfer outcomes.

EmbryoGlue® (Vitrolife) is a hyaluronan-rich medium resembling uterine fluid. It is reported to facilitate rapid diffusion with viscous uterine fluid, provide nutritional support from embryo transfer to implantation and support biochemical signaling during implantation (Vitrolife Fertility Systems, 2004). The effects of EmbryoGlue® used during in vitro fertilization − embryo transfer (IVF-ET) in our laboratory is evaluated. Retrospective study. 1633 IVF-ET cases were included from 01/01/2002 to 3/15/2004 at ART Reproductive Center, Beverly Hills, California. During stage one (S1) modified HTF-hepes (Irvine Scientific, Cat 90126) + 20%SS (Irvine Scientific, Cat 90193) was used for ET. Stage 2 (S2) used EmbryoGlue® as the transfer medium according to the manufacturer’s suggested guidelines. In comparing the overall clinical pregnancy rates (Day 3--Day 6 transfers), with EmbryoGlue® group was 39.75% (221/556) which is significant lower as compared to 45.87% (494/1077) in the without EmbryoGlue® group (p<0.05). Compare the Day 3 transfer result, 35.84% (119/332) of the EmbryoGlue® group versus 45.28% (321/709) of the without EmbryoGlue® group with significant difference (p<0.05). Evaluation of the blastocyst transfer (Days 5–6 post egg retrieval) rate was 61.18% (85/544) in the “EmbryoGlue®” group versus 55.49% of the “without EmbryoGlue®” group. While the pregnancy rate is higher, the difference was not significant. Our data demonstrate that EmbryoGlue® group has low pregnancy rate in day 3 transfers. During this study, the culture media changed from the “G-II” series used in the Non-Embryo Glue® group to the GIII series, which was used for the Embryo Glue® group. Presumably this did not account for the lack of difference observed between both arms of the study. We speculate that the advantages of Embryo Glue® may be pronounced in the later stages of embryo development when receptors and biochemical signaling between the blastocyst and the uterus may benefit from its presence in the milieu at this critical stage. üView Large Image Figure ViewerDownload Hi-res image Download (PPT)

This study involved examining the relative percentages of male and female embryos from the Preimplantation Genetic Diagnosis (PGD) analysis of fertile patients undergoing sex selection for a history of multiple births of a single sex. The goal was to determine if such couples produce embryos of predominantly one sex, thus predisposing them to having same-sex children. A retrospective chart-review study. All PGD summaries from our center from January 2003 to April 2004 were reviewed (n=332). Patient charts were requested for those summaries that quoted “sex selection” as an indication for PGD, as well as those summaries that recorded embryo transfers of only one sex. 80 such charts were then analyzed for patients requesting PGD for sex selection based on a history of bearing three or more children of the same sex. All births of the current couple and any previous offspring of the male partner were included. Exclusion criteria included fewer than three same-sex children, a new male partner, donor sperm, and/or a diagnosis requiring ICSI (a procedure which may bypass natural selection bias for sperm). Couples with five or more same-sex children were not excluded for having one offspring of the minority sex. Nine couples met criteria, having 33 of 35 children among them of a single sex (94.3%). Four couples had males and five couples had females. Thirty-one of their combined 33 pregnancies (93.9%) were spontaneous (both twin pregnancies were IVF). All embryos biopsied for PGD from these couples were tallied. A total of 60 embryos were biopsied from the 9 couples meeting study criteria over 10 cycles (one couple requested PGD for two cycles). Thirty-four embryos (56.7%) were identified as the desired sex (opposite the sex already achieved), compared with 24 embryos (40%) of the non-desired sex. The desired sex outnumbered the previously achieved sex in both normal (20 vs.16) and abnormal (14 vs. 8) embryos. The female-desired group conceived six normal males and 13 normal females. The male-desired group conceived 7 normal males and 10 normal females. Only two embryos (3.3%) could not be assigned a sex. In this study, couples known to produce offspring of predominantly one sex actually produced more of the opposite sex when all embryos were counted together using PGD for sex determination. Three couples did have sperm separation for sex selection prior to insemination, but in only one instance did this result in more embryos of the desired sex (less than would be expected via chance alone). The resulting conclusion is that ordinary chance led to the same-sex children, as opposed to a predisposition to having a particular sex. This study may not account for in-vivo selection bias of sperm or embryos. It does, however, suggest that such couples are able to conceive offspring of both sexes.