Livebirth derived from in vitro matured oocytes.

The authors presented a case report of a woman who suffered from primary infertility due to anovulation. She was diagnosed to have polycystic ovarian syndrome (PCOS) and was treated by ovarian stimulation with conventional technique, such as clomiphene citrate alone, the combination of clomiphene citrate and metformin and the combination of clomiphene citrate with gonadotropins. Nevertheless all of them resulted in no response. Therefore, in vitro maturation of oocyte (IVM) was started and followed by intracytoplasmic sperm injection (ICSI) of 22 oocytes. 14 embryos developed and three of them were transferred back to the uterine cavity. She conceived and the ultrasonography revealed twin pregnancy at the 8-weeks gestation. She gave birth at the 31 weeks’ gestation in Siriraj Hospital. The newborns were admitted in the hospital for 24 days for phototherapy in treating jaundice which was due to prematurity. Then both of them were discharged healthily.

A comparison between conventional slow freezing and vitrification of mouse blastocysts using cryo-E.

Objective: To compare the efficiency of two cryopreservations between conventional slow freezing and vitrification of mouse blastocysts using cryo-E. Methods: ICR female mice (8 weeks) were superovulated with 5 IU/ml of pregnant mare serum gonadotrophin (PMSG), the successfulness of mating with males was verified by the presence of a vaginal plug. Blastocysts were obtained between 3.5 and 4.5 days per p.c. or 96-108 hours after hCG administration by flushing the uterus. Randomly selected blastocysts were simultaneously frozen by slow-rate freezing and a vitrification method. One month later, the embryos were thawed and cultured in the blastocyst medium (COOK; Sydney IVF, Australia). Results: Based on 250 slow freezing and 310 vitrified mouse blastocysts, vitrification resulted in a slightly higher survival and hatching rates than the slow-freezing method (83.9% VS 82.0%, and 68.8% VS 66.8%, respectively). Conclusion: Both slow freezing and vitrification of mouse blastocysts are useful methods for cryopreservation. These results showed that vitrification is better than slow freezing in terms of simplicity, duration, and cost-effectiveness.

Preliminary data of preimplantation genetic diagnosis for aneuploidy screening using fluorescent in situ hybridization in Siriraj Hospital.

Objective: Preimplantation genetic diagnosis (PGD) is technique for detecting genetic diseases. Preimplantation genetic diagnosis for aneuploidy screening (PGD-AS) using fluorescent in situ hybridization (FISH) has been used worldwide including at Siriraj Hospital. The objective of this study was to comparethe pregnancy rate between a PGD-AS group with standard assisted reproductive techniques (ARTs) in Siriraj Hospital. Methods: Couples who requested PGD-AS underwent a standard ARTs process followed with blastomere biopsy for FISH analysis. The pregnancy rate was compared among the PGD-AS group and the control group. The control group was divided into 2 subgroups – all patients required ARTs subgroup and age ≥ 35 yrs. subgroup. Results: 6 stimulated cycles from 4 patients were performed in the PGD-AS group. The pregnancy rate per stimulated cycle in the PGD-AS group, control group and age ≥ 35 yrs group were 33.33%, 16.20% and 12.05% respectively. Moreover, the pregnancy rate per transferred cycles in the PGD-AS group, control group and age ≥ 35 yrs group were 40.00%, 21.02% and 13.17% respectively. Conclusion: PGD is an advanced method for detecting genetic defects. PGA-AS might increase the pregnancy rate.

Correlation between hyaluronan binding assay and TUNEL assay for sperm DNA integrity identification.

Objective: To study the correlation between the percentages of HA-unbound sperm and DNA fragmented sperm by TUNEL assay. Methods: The semen residue from semen analysis was tested by HBA and TUNEL assay. Results: The mean age of patients included in the study was 34.8 years (± 3.7 years). The proportion of HA-unbound sperm ranged from 11.3% to 24.2%, with a mean of 17.08% (± 3.24%). The range of TUNEL positive in semen samples was 2% to 11.75%, with a mean of 5.78% (± 2.28%). Pearson’s correlation between two tests was 0.848 (p<0.01). Intraobserver variation of the results of HBA ranged from 3.3% to 7.6%, with a mean of 6.23% (± 1.11%). Intraobserver variation of the results of TUNEL assay ranged from 0% to 6.9%, with a mean of 1.54% (± 2.7%). Agreement measuring of each test was determined by using intraclass correlation. The intraclass correlation coefficient of HBA and TUNEL assay were 0.970 (P<0.001) and 0.997 (P<0.001) respectively. Conclusion: As several studies have found, the binding capacity of sperm to HA is correlated with several sperm parameters. In this study, the strong correlation between the percentages of HA-unbound sperm and TUNEL positive sperm implies, furthermore, that the HA-bound sperm percentage correlates with low levels of DNA fragmentation.

Vitrification of mouse oocyte using open pulled straws compared with needles.

OBJECTIVE: To compare the survival rate of mouse oocytes and fertilization rate between using open pulled straws (OPS) and needles for vitrification. MATERIAL AND METHOD: Meiosis II oocytes from female C57B/6J mice aged 7-8 weeks were collected and allocated to two groups for vitrification by using OPS or needles. Vitrified oocytes were thawed, morphological survival and fertilization rate were examined. RESULTS: There was no obvious difference between the morphological survival rates of vitrified mouse oocytes using OPS and needles (66.7% vs 64.8%). Proportions Difference 1.9% (95% CI -7.1, 10.7). The vitrified oocytes from the needle had significantly higher percentages of fertilization rate than OPS (76.8% vs 62.5%). Proportions Difference -14.3% (95% CI -24.5, -3.6). CONCLUSION: Vitrification method of mouse oocytes using needles when compared to OPS provides a similar morphological survival rate and higher fertilization rate.