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'Curing' empty follicle syndrome.

We report a novel method of rescuing empty follicle syndrome (EFS) and provide evidence that it is a drug-related problem rather than a clinical dysfunction. In a preliminary study we established that in EFS the serum beta-human chorionic gonadotrophin (beta-HCG) concentrations 36 h after HCG administration never exceeded 10 mIU/ ml. beta-HCG concentrations were thus used to confirm EFS when oocytes were not retrieved from one ovary after controlled ovarian hyperstimulation. The procedure was suspended leaving intact all follicles in the second, ovary. After confirmation of EFS, a second HCG from a different batch was administered and 36 h later mature oocytes were retrieved from the intact ovary, suggesting a fault with the HCG previously administered. Three patients have been treated in this way. In the first case, four out of five mature eggs were fertilized after intracytoplasmic sperm injection (ICSI) resulting in the transfer of three top grade (grade 1) embryos. In the second case all seven mature oocytes fertilized after in-vitro fertilization (IVF) and three grade 1 embryos were transferred resulting in a twin pregnancy, now delivered. In the third case, five out of nine oocytes were fertilized after ICSI and one out of the three treated with high insemination concentration IVF fertilized, resulting in the transfer of three ICSI embryos.

Chorionic Gonadotropin↗

Development and implementation of intracytoplasmic sperm injection (ICSI).

The purpose of this paper is to elucidate the experimental steps that led to the development of intracytoplasmic sperm injection (ICSI) and its application in the human. ICSI has become the most successful micromanipulation procedure for treating male infertility. A total of 355 in vitro fertilization (IVF) cycles utilizing ICSI are described; 180 couples were previously treated in 509 IVF cycles but achieved no fertilization and 175 couples could not be treated by IVF because of extremely poor semen parameters. Of the 3063 metaphase II (M II) oocytes retrieved, 2970 were injected with a survival rate of 93.6%, yielding 1917 bipronuclear zygotes (64.5%). In 148 patients, a foetal heart was evidenced by ultrasound; 11 of these patients miscarried between 7 and 13 weeks of gestation. The ongoing pregnancy rate was 38.6% (137/355) per retrieval and 40.5% (137/338) per embryo replacement. At the time of writing, there were 22 deliveries and one therapeutic abortion for a trisomy 21 chromosomal abnormality. In addition, 66 singleton, 37 twin, 10 triplet and 1 quadruplet pregnancies were ongoing. The concentration of motile spermatozoa in the ejaculate only slightly influenced the fertilization rate (P < 0.001) and the pregnancy outcome (P < 0.01). A preliminary injection procedure utilizing intracytoplasmic injection of isolated sperm heads was performed in 35 M II human oocytes with resultant fertilization and cleavage rates of 74% and 73% respectively. Skills in ICSI were acquired by injecting hamster and unfertilized human oocytes with human sperm. ICSI can be used to successfully treat couples who have failed IVF or who have too few spermatozoa for conventional in vitro insemination.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The acrosome index, radical buffer capacity and number of isolated progressively motile spermatozoa predict IVF results.

BACKGROUND: The accuracy by which a number of newly described semen variables can predict either total fertilization failure (TFF) or pregnancy outcome in IVF, has not previously been investigated. The study aim was, therefore, to determine prospectively the predictive value of these variables. METHODS: The semen variables investigated were the post-wash total progressively motile sperm cell count (TPMC(post-wash)), the acrosome index (AI), 'cytoplasmic residues' and normal sperm morphology, evaluated according to the strict criteria ('strict criteria'), as well as the fast and slow total radical trapping antioxidant potential ('fast TRAP' and 'slow TRAP' respectively). RESULTS: The study group (n = 87) showed a mean (+/- SD) number of 10.2 +/- SD retrieved oocytes, 12.6% TFF, a mean fertilization rate of 59.7% and a pregnancy rate of 19.5% (17/87). TFF was significantly predicted by TPMC(post-wash), 'strict criteria', AI and 'cytoplasmic residues' (all P < 0.05). The outcome after embryo transfer was significantly predicted by AI and 'fast TRAP'. Semen samples with an AI <5% and a 'fast TRAP' <1.14 mmol/l in particular did not result in any pregnancies after IVF-embryo transfer. CONCLUSIONS: Of all the measured and calculated semen variables, TPMC(post-wash) was the best predictor of TFF, whilst AI and 'fast TRAP' were the best predictors of pregnancy after IVF.

Acrosome Reaction↗

The clinical efficacy and efficiency of a 100-IU starting dose of recombinant follicle stimulating hormone (Puregon) in Korean women undergoing in vitro fertilization and embryo transfer.

AIM: A prospective, non-comparative study was conducted to evaluate the efficacy and efficiency of a starting dose of 100 IU recombinant follicle stimulating hormone (rhFSH, Puregon) in women undergoing ovarian stimulation prior to in vitro fertilization (IVF). METHODS: A total of 40 women were down-regulated with gonadotropin releasing hormone agonist (long protocol), then treated with a fixed dose of 100 IU rhFSH for the first 4 days, and the dose of gonadotropin administration was adjusted according to patient's response thereafter. A maximum number of 3 embryos were transferred in 32 patients. RESULTS: Nine patients were treated with intracytoplasmic sperm injection, and 24 with conventional IVF. The duration of stimulation was 10.8 +/- 1.8 days, serum estradiol concentration on human chorionic gonadotropin day was 1693.0 +/- 1651.2 pg/mL, and 1480 +/- 450 IU rhFSH was used. A mean number of 8.4 +/- 5.7 oocytes were retrieved, 4.9 +/- 4.3 embryos obtained, 2.4 +/- 0.9 embryos transferred, and 3.5 +/- 3.2 embryos cryopreserved. The proportion of mature oocytes was 71.6%, and the fertilization rate was 86.4%. Clinical pregnancy was achieved in 8 patients (25.0%), and all of these pregnancies are ongoing or delivered. There were 3 cases of twin gestations (37.5%). The implantation rate was 13.1% (11/84). There was one case of moderate ovarian hyperstimulation syndrome, however, the patient recovered within 7 days without any complications. CONCLUSION: The starting dose of 100 IU rhFSH has a good safety profile, and is adequate in controlled ovarian hyperstimulation for IVF with a small amount of gonadotropin administered.

Adult↗

Male factors determining the outcome of intracytoplasmic sperm injection with epididymal and testicular spermatozoa.

During a period of 8 years, 1,079 intracytoplasmic sperm injection (ICSI) procedures with aspirated epididymal or testicular spermatozoa were performed. Epididymal spermatozoa were used in 172 cycles and testicular spermatozoa or spermatids in 907 cycles. Multiple biopsies were obtained from at least two different locations in the testes. Retrieved spermatozoa were used after cryopreservation (frozen) or immediately after aspiration (fresh). Three hundred patients had obstructive azoospermia (OA) or ejaculation failure. In 414 cases, azoospermia was caused by impaired spermatogenesis resulting from maldescended testes, chemotherapy/radiotherapy, or by Sertoli-cell-only syndrome, genetic disorders or unknown aetiology. Transfer rates, pregnancy rates and birth rates per ICSI cycle showed no statistically significant differences between testicular and epididymal spermatozoa in men with OA (28% average birth rates in both cases). However, birth rates differed significantly with regard to the status of spermatogenesis. Treatment of men with nonobstructive azoospermia (NOA) resulted in a birth rate of 19% per cycle. In all patient groups, there was no difference in the birth rates achieved with fresh and cryopreserved spermatozoa. While testicular volume, follicle-stimulating hormone level and age of the male patient are no statistically significant prognostic factors, the underlying cause of azoospermia is the most important factor determining the outcome of ICSI with epididymal and testicular spermatozoa. The pregnancy rate is lower in NOA patients than in those with OA.

Epididymis↗

Fertilization, pregnancy and embryo implantation rates after ICSI in cases of obstructive and non-obstructive azoospermia.

The aetiology of azoospermia can be grossly divided into obstructive and non-obstructive causes. Although in both cases testicular spermatozoa can be used to treat male fertility, it is not well established whether success rates following intracytoplasmic sperm injection (ICSI) are comparable. Therefore, a retrospective analysis of fertilization, pregnancy and embryo implantation rates was performed following ICSI with testicular spermatozoa in obstructive or non-obstructive azoospermia. In total, 193 ICSI cycles were carried out with freshly retrieved testicular spermatozoa; in 139 cases of obstructive and 54 cases of non-obstructive azoospermia. The fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia was significantly lower than in obstructive azoospermia (67.8% versus 74.5%; P = 0.0167). Within the non-obstructive group, the fertilization rate in the group of maturation arrest (47.0%) was significantly lower than in case of Sertoli cell-only (SCO) syndrome (71.2%) or germ cell hypoplasia (79. 5%). Embryo quality on day 2 after ICSI was similar for all groups. Pregnancy rates per transfer between obstructive (36.8%) and non-obstructive groups (36.7%) were similar. In cases of maturation arrest the pregnancy rate per transfer was lowest (20.0%) although not significantly different from SCO syndrome or hypoplasia groups. Embryo implantation rates were not different between the obstructive (19.6%) and non-obstructive groups (25.8%), and were lowest in cases of germ cell hypoplasia (15.8%). This retrospective analysis shows that although fertilization rate after ICSI with testicular spermatozoa in non-obstructive azoospermia is significantly lower than in obstructive azoospermia, pregnancy and embryo implantation rates are similar.

Adult↗

Predictive factors for motile sperm recovery using testicular biopsy in nonobstructive azoospermic patients.

OBJECTIVE: The objective of the present study was to assess the predictive value of hormone values, histological analysis for the finding of motile spermatozoa on testicular biopsy in nonobstructive azoospermia. METHODS: This cross-sectional study assessed serum follicle-stimulating hormone (FSH), luteinizing hormone, prolactin and total testosterone values in 50 patients undergoing testicular biopsy prior to ICSI. P < 0.05 was considered significant. RESULTS: Recovery was successful in 28 cases, and motile spermatozoa were retrieved in 7. Significant differences were detected between presence of motile spermatozoa and absence of spermatozoa in terms of FSH values (P = 0.003, one-way ANOVA). The other variables did not present statistical differences. A receiver operating characteristic curve showed that FSH levels below 17.00 IU/L were predictive of motile spermatozoa recovery. CONCLUSIONS: FSH values can serve as a predictive factor for the recovery of motile spermatozoa using biopsy in azoospermic patients. On the other hand, histological analysis and other hormone values were not helpful in the prediction of motile sperm.

Biopsy↗

Distinction between paternal and maternal contributions to the tripronucleus in human zygotes obtained after in vitro fertilization.

OBJECTIVE: The aim of this study was to determine the genetic composition of tripronuclear (3PN) zygotes. Fluorescence in situ hybridization (FISH) was used to quantify the sperm tails in human 3PN zygotes. MATERIALS AND METHODS: Ovarian hyperstimulation was performed using a standard long protocol consisting of gonadotropin-releasing hormone agonist (GnRHa) with human menopausal gonadotropins (HMG) and/or recombinant follicle stimulating hormone (rFSH). Human chorionic gonadotropin (hCG) was administered when at least two leading follicles reached 18 mm in mean diameter. Oocytes were retrieved from the follicles transvaginally under ultrasound guidance 34-36 hours after administration of hCG. The oocytes were examined 16-18 hours after fertilization for the presence and number of pronuclei. The 3PN zygotes were then fixed and extracted in buffer at 37 degrees C for 60 minutes. After washing in a blocking solution, the 3PN zygotes were prepared for indirect immunofluorescence using monoclonal antibody and Hoechst dye 33342 to visualize the number of pronuclei. RESULTS: Twenty-one 3PN zygotes were randomly collected for evaluation. The number of 3PN zygotes containing one, two or four sperm tails were two (10%), 18 (85%) and one (5%), respectively. CONCLUSION: Based on our observations, additional pronuclei are mainly of paternal origin.

Cell Nucleus↗

Comparison of a gonadotropin-releasing hormone (GnRH) antagonist and GnRH agonist flare-up regimen in poor responders undergoing ovarian stimulation.

OBJECTIVE: To compare the efficacy of flare-up and GnRH-antagonist treatment in poor-responder patients. DESIGN: Randomized prospective study. SETTING: Assisted reproduction center. PATIENT(S): Fifty-five poor-responder patients undergoing intracytoplasmic sperm injection (ICSI). INTERVENTION(S): Thirty patients received GnRH agonist on the 1st day of menstruation, followed by exogenous gonadotropins from the 2nd day. Twenty-five patients received exogenous gonadotropins starting on the second day of menstruation, followed by GnRH antagonist when the leading follicle reached 14 mm in diameter. MAIN OUTCOME MEASURE(S): The total dose of FSH administered during the ovarian stimulation, as well as the number of mature oocytes retrieved, embryo quality, fertilization, implantation, and pregnancy rates were evaluated. RESULT(S): The number of ampules and units of FSH administered were significantly less in the flare-up than in the antagonistic group. The numbers of mature oocytes retrieved and of top-quality embryos transferred were significantly greater in the flare-up than in the GnRH-antagonist group. The fertilization rate (84% vs. 63%) was significantly higher in the flare-up than in the GnRH-antagonist group. The implantation and pregnancy rate were similar in the two groups. CONCLUSION(S): The flare-up protocol appears to be more effective than the GnRH-antagonist protocol in terms of mature oocytes retrieved, fertilization rate, and top-quality embryos transferred in poor-responder patients.

Adult↗

Efficacy of the sperm survival test for the prediction of oocyte fertilization in culture.

The present study was carried out to investigate the predictive value of the sperm survival test (SST) with respect to the fertilization of oocytes in culture. In general, our laboratory uses a total of 50,000-150,000 motile spermatozoa to inseminate each oocyte. The remaining material is evaluated for motility before and after 24 h of incubation at 37 degrees C in a 5% CO2 atmosphere. A total of 250 oocytes from 50 cases (mean +/- SD, 5.0 +/- 2.4 oocytes per retrieval) were inseminated and the final rate of cleaved embryos obtained was 52.5%. The SST (%) was considered normal when the ratio (final density of progressing spermatozoa after 24 h x 100/initial density of progressing spermatozoa) was 50% or more. Any other result was considered abnormal. Cases presenting one or more cleaved embryos (n = 40) were separated from those in which no embryo formation occurred (n = 10) and the results were compared in terms of the respective sperm survival rates over a period of 24 h: normal SST (one or more cleaved embryos, 37; none, five), abnormal SST (one or more cleaved embryos, three; none, five). The specificity of the SST was 0.92 and sensitivity 0.50, the predictive value of the abnormal test was 0.62 and the predictive value of the normal test 0.88. The efficacy of the test was estimated at 0.71, which was better than the conventional parameters of sperm analysis. A receiver-operating characteristics curve for SST confirmed that the test can be useful for the prediction of fertilizability of oocytes in the laboratory.

Adult↗

Intracytoplasmic injection with late spermatids: a successful procedure in achieving childbirth for couples in which the male partner suffers from azoospermia due to deficient spermatogenesis.

OBJECTIVE: To investigate the feasibility of using late spermatids in intracytoplasmic sperm injection (ICSI) for patients in which there is a total lack of normal and testicular sperm. DESIGN: Clinical study. SETTING: Private fertility center with adjacent laboratory facilities. PATIENT(S): Thirty-six males diagnosed with azoospermia underwent testicular biopsy and the resulting spermatids from nine patients were used to fertilize oocytes retrieved from their respective wives by ICSI. INTERVENTION(S): Intracytoplasmic spermatid injection of oocytes with late spermatids obtained by testicular biopsy. MAIN OUTCOME MEASURE(S): Fertilization, pregnancy, and delivery rates. RESULT(S): Three of the cases resulted in pregnancies with childbirth occurring in all three, identical twins in one, and individual singletons in the other two cases. CONCLUSION(S): It is possible to achieve successful levels of pregnancy and childbirth in cases of azoospermia by intracytoplasmic injection of late spermatids.

Birth Weight↗

[Optimisation of possible success in an IVF program].

The success of an IVF (in vitro fertilisation) programme is mainly dependent an the quality of the IVF laboratory. Beside this the quality of ovarian stimulation play another, but not as important role. In the 4 (th) quarter of 2003 we have changed the culture media for oocytes and embryos in the IVF laboratory of ENDOKRINOLOGIKUM Hamburg from simple to complex media (G-1 version 3). As protein source we used HSA-solution. Before embryo transfer equilibration for 1 hour in EmbryoGlue was performed, which contains hyaluran. Hyaluran is said to improve implantation rates by different ways. The media were bought from Vitrolife Schweden AB (Kungsbacka, Schweden) via a German distributor. These data were compared to those of two previous 3-month-periods (3rd quarter 2003 und 4th quarter 2002). The study design was prospective, with two historical control groups. In the three study periods 255, 343 and 503 patients were treated by either conventional IVF or ICSI (intracytoplasmic sperm injection). The cohorts were comparable regarding their anamnestic data. The number of patients treated which the GnRH antagonist cetrorelix (Cetrotide) significantly increased from 24.9 % to 49.5 % and 60.9 %. Under this protocol significantly less oocytes were retrieved (10.23 vs. 8.73 vs. 8.88), leading to slightly less embryos (1.94 vs. 1.91 vs. 1.85, not significant, n. s.). The cumulative embryo score significantly decreased from 28.58 to 24.10. It was significantly higher in the study cohort as compared to the two control cohorts (32.44). The clinical pregnancy rate significantly increased to 25.29 % in the study cohort as compared to the two historical control cohorts (15.02 % and 18.75 %). We could achieve a significant improvement in the success rates of our IVF program by switch from simple to more complex culture media in the IVF laboratory. The use of the GnRH antagonist protocol with cetrorelix (Cetrotide) lead to more comfort to the patients but did not influence the results.

Adult↗

Reproductive potential of fresh and cryopreserved epididymal and testicular spermatozoa in consecutive intracytoplasmic sperm injection cycles in the same patients.

OBJECTIVE: To determine if the cryopreservation of epididymal and testicular spermatozoa alters their reproductive potential by examination of patients who underwent consecutive cycles of ICSI using fresh and then cryopreserved spermatozoa. DESIGN: Retrospective review. SETTING: Tertiary care university hospital. PATIENT(S): One hundred sixty-two consecutive cycles of ICSI were analyzed. Thirteen patients were identified as having undergone treatment with freshly retrieved epididymal spermatozoa; these patients subsequently underwent treatment with spermatozoa cryopreserved from that cycle. Eighteen patients underwent ICSI with freshly retrieved testicular spermatozoa; these patients subsequently underwent treatment with spermatozoa cryopreserved from that cycle. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Fertilization rates and pregnancy rates. RESULT(S): The fertilizing capacity of epididymal spermatozoa remained unchanged after cryopreservation and subsequent thawing, with fertilization rates of 58% and 57% for fresh and cryopreserved spermatozoa, respectively. Testicular spermatozoa, however, showed a significant decrease in fertilizing capacity after cryopreservation when compared with freshly retrieved spermatozoa (52% and 71%, respectively). Pregnancy rates appeared unaffected by the cryopreservation of epididymal spermatozoa (fresh, 3/13; frozen, 2/13) or testicular spermatozoa (fresh, 2/18; frozen, 5/18). CONCLUSION(S): This study offers further evidence that motile epididymal spermatozoa retain their fertilizing capacity after cryopreservation. The data presented on testicular spermatozoa suggest that although cryopreservation may reduce the fertilizing capacity of testicular spermatozoa, there is no decrease in pregnancy rates.

Adult↗

The role of purified follicle stimulating hormone therapy in the male partner before intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the impact of long-term purified FSH (pFSH) therapy in male partners before intracytoplasmic sperm injection (ICSI). DESIGN: Prospective, randomized, controlled study. SETTING: Large university-based IVF unit. PATIENT(S): Seventy-eight patients made up the study and control groups (39 patients each). All patients had severe male factor infertility. INTERVENTION(S): Induction of ovulation, oocyte retrieval, ICSI, and ET were carried out in both groups. In the study group, male partners received pFSH (75 IU FSH, <1 IU LH) for > or = 50 days before oocyte retrieval. MAIN OUTCOME MEASURE(S): Fertilization rate, embryo quality, implantation and pregnancy rates. RESULT(S): Fertilization and pregnancy rates were higher in the study group (68% and 35.9%, respectively) were higher than in the control group (59% and 17.9%, respectively), although the differences did not reach statistical significance. The implantation rate was significantly higher in the study group (15.5% versus 6.5%). The study group showed a trend toward a higher number of better-quality embryos per transfer (mean +/- SD, 2.2 +/- 1.6 versus 1.6 +/- 1.6). CONCLUSION(S): Purified FSH therapy in male partners before ICSI improves implantation rate. Improved embryo quality may be a contributory factor.

Embryo Implantation↗

Assisted reproductive technologies (ART) in Canada: 2001 results from the Canadian ART Register.

OBJECTIVE: To present the success rates of assisted reproductive technologies (ART) cycles performed in 2001 in Canada. DESIGN: Retrospective cohort study. SETTING: Nineteen of 22 ART centers in Canada. PARTICIPANTS: Couples undergoing ART treatment in Canada during 2001. METHODS: Data on each ART cycle performed during 2001 were submitted electronically to the Canadian ART Register (CARTR) by participating centers. MAIN OUTCOME MEASURE(S): Clinical pregnancy and live birth rate per cycle started, multiple birth rate. RESULT(S): A total of 7,884 ART cycles was reported to CARTR. There were 5,393 in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles using the woman's own oocytes. Per cycle started, the pregnancy rate was 28.3%, and the live birth rate was 23.1%; the multiple birth rate per delivery was 32.8%. Of cycles with oocytes retrieved, IVF was performed in 44% and ICSI in 56%; the outcomes were similar with the two procedures. There were 301 IVF/ICSI cycles using donor oocytes. The pregnancy rate was 29.2%, and the live birth rate was 22.4%; the multiple birth rate was 43.5%. There were 1,936 frozen embryo transfer cycles using the woman's own oocytes. The pregnancy rate was 18.9%, and the live birth rate was 15.4%; the multiple birth rate was 24.9%. CONCLUSION(S): For 2001, CARTR achieved 86% voluntary participation from Canadian ART centers. Pregnancy and live birth rates comparable to those of other countries were achieved.

Adult↗

Testicular tissue extraction in a young male with 47,XXY Klinefelter's syndrome: potential strategy for preservation of fertility.

OBJECTIVE: To describe a case involving the cryopreservation of testis tissue retrieved from a 15-year-old male teenager with Klinefelter's syndrome. DESIGN: Case report. SETTING: An academic medical center. PATIENT(S): A 15-year-old boy with Klinefelter's syndrome. INTERVENTION(S): Microsurgical testis sperm extraction with cryopreservation of harvested tissue. MAIN OUTCOME MEASURE(S): Spermatozoa within testis tissue. RESULT(S): Successful extraction and cryopreservation of three vials of sperm-containing testis tissue. No effect on subsequent testosterone levels. CONCLUSION(S): Testis tissue extraction in the adequately virilized but azoospermic young male with 47, XXY Klinefelter's syndrome may be a strategy to preserve future biological paternity.

Adolescent↗

Successful in vitro maturation of human oocytes not exposed to human chorionic gonadotropin during ovulation induction, resulting in pregnancy.

OBJECTIVE: To report a case of successful in vitro maturation of human oocytes not exposed to hCG during ovulation induction, resulting in pregnancy after transfer of a frozen-thawed embryo resulting from intracytoplasmic sperm injection (ICSI) of the in vitro-matured human oocytes. DESIGN: Clinical study. SETTING: Hospital-based private IVF center. PATIENT(S): A couple with infertility due to severe oligospermia. INTERVENTION(S): Five immature oocytes were retrieved from a patient who failed to use hCG during ovarian stimulation and were cultured for 48 hours in B2 medium containing FSH and hCG. Five oocytes extruded the first polar body and underwent ICSI with frozen-thawed husband spermatozoa. MAIN OUTCOME MEASURE(S): In vitro maturation, fertilization after ICSI, embryo development, and pregnancy. RESULT(S): All five oocytes extruded the first polar body and were injected using ICSI. Three oocytes were fertilized, but two showed three pronuclei. The remainder, a two-pronuclei embryo subsequently cleaved and was cryopreserved. An ongoing pregnancy was obtained after-the transfer of this frozen-thawed embryo. CONCLUSION(S): Immature human oocytes at the germinal-vesicle stage that have not been exposed to hCG during ovarian stimulation can be matured in vitro and a normal pregnancy can result from ICSI of the in vitro-matured oocytes.

Adult↗

Preimplantation diagnosis for X and Y normality in embryos from three Klinefelter patients.

In some 47,XXY Klinefelter patients without evidence of mosaicism, testicular spermatozoa can be successfully recovered and used for intracytoplasmic sperm injection (ICSI). To ensure the replacement of embryos with a normal X and Y chromosome pattern, preimplantation diagnosis can be performed. This paper reports on three 47,XXY Klinefelter patients from whom it was possible to retrieve testicular spermatozoa in order to perform ICSI. From their healthy wives a total of 27 oocyte-cumulus complexes were retrieved from which 22 metaphase-II oocytes were obtained and injected; 19 of these were intact after injection. Two distinct pronuclei were observed in eight oocytes (42.1%) 18 h after injection. On day 3 of development, five embryos (62.5%) had reached at least the 6-cell stage and were of sufficient quality to undergo biopsy and subsequent preimplantation diagnosis for sex chromosome analysis by fluorescent in-situ hybridization. The four embryos diagnosed as normal were transferred to three respective patients, resulting in one biochemical pregnancy. The remaining cells of the fifth embryo were analysed afterwards, revealing a normal X and Y chromosome constitution. So far, in the five embryos diagnosed, a normal sex chromosome pattern has been observed.

Embryo, Mammalian↗