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.

Heterotopic pregnancy following tuboplasty and ovulation induction by clomiphene citrate and timed intrauterine insemination: Case report.

A Thai woman 36 year old with post tubal sterilization. She was performed for Tuboplasty to restore fertility on new marriage. Both fallopian tubes functioned post correction. She tried natural conception, but was not successful. The doctor treated her by the way of ovarian induction with Clomiphene Citrate and intrauterine insemination. She conceived in the 4th cycle, but she had an heterotopic pregnancy. Gestational sac, fetal echo and positive fetal heart beat were detected at both the intrauterine cavity and the right adnexa, and she had no abnormal symptoms. The Diagnostic Laparoscopy was performed. Following the operation an unruptured right tubal pregnancy was found. Right salpingectomy was done and the pathological report confirmed tubal pregnancy. Post operative time, she had no signs of abortion, and normal intrauterine growth was detected.

Freezing effect on post-thawed sperm characteristics especially sperm DNA integrity comparing between liquid nitrogen vapour and computerized program freezer.

The aim of the study was to evaluate the cryodamage effects on human sperm characteristics, especially on sperm DNA integrity, after 6 months of freezing comparing between liquid nitrogen vapour (LNV) and computerized program freezer (CPF). Forty normal semen samples were collected for semen analysis. Each sample was mixed with cryoprotective media and devided into 2 straws. The first straw was frozen with LNV and the second one with CPF. After 6 months of cryostorage, semen samples were thawed, and sperm chromatin integrity as well as sperm motility, morphology, vitality and cryosurvival rate were determined. Percentages of DNA damage were higher (p<0.01) following freezing with LNV than with CPF. Sperm vitality was greater (p<0.05) after CPF than after LNV, as well as cryosurvival rate (p<0.001). Post-thawed sperm motility was greater after CPF than after LNV, either in grade A (p<0.001) or in grade B (p<0.05). No significant difference was observed in the percentage of normal sperm morphology comparing the two freezing methods. The current study demonstrated post-thawed decrease in sperm DNA integrity as well as other sperm characteristics after freezing in both methods. The CPF significantly provided superior results in post-thawed sperm DNA integrity, sperm motility and vitality than LNV did. In case of 6 months of cryostorage, therefore, we recommend the computerized program freezer as a preference for sperm cryopreservation.

Comparison between sperm treated with pentoxifylline and 2-deoxyadenosine using hypo-osmotic swelling test.

OBJECTIVE: To compare the percentage of sperm tail membrane swelling under hypo-osmotic conditions between sperm treated with pentoxifylline and 2-deoxyadenosine. DESIGN: Experimental in vitro study. MATERIAL AND METHOD: Thirty normal semen samples from male partners of infertile couples were collected. After sperm preparation by two-layer Percoll gradient method, each sperm sample was divided into three specimens. Pentoxifylline and 2-deoxyadenosine were separately added into two specimens, while the third specimen was used as a control. Hypo-osmotic swelling test was performed in all specimens. Percentage of swollen spermatozoa in each specimen was evaluated. RESULTS: The mean percentage of swollen spermatozoa in the semen samples supplemented with pentoxifylline and 2-deoxyadenosine were both significantly higher than those in the control (82.8 +/- 7.7 and 83.0 +/- 9.5 vs 70.8 +/- 12.7; p < 0.001). There was no significant differences of swollen spermatozoa between pentoxifylline and 2-deoxyadenosine (p = 0.898). CONCLUSION: Addition of pentoxifylline and 2-deoxyadenosine to the sperm prepared by the two-layer Percoll gradient method can almost equally enhance the sperm membrane integrity. Therefore, it may be beneficial to add these compounds to sperm preparation for use in assisted reproduction.

Hypoosmotic swelling test of sperm supplemented with pentoxifylline after preparation by two-layer percoll gradient method.

The aim of this study was to compare the percentage of sperm tail membrane swelling under hypoosmotic conditions with those with and without pentoxifylline supplements in sperm prepared by the two-layer Percoll gradient method. Twenty five normal semen samples were collected from male partners of infertile couples attending the Infretility Clinic at Siriraj Hospital. After the process of sperm preparation by the two-layer Percoll gradient method, the final samples were divided into 2 tubes, 0.5 ml was added into one tube and another tube was kept as control. The hypoosmotic swelling test was performed on both specimens. The percentage of swollen spermatozoa in the pentoxifylline supplement group was significantly higher than the control group (82.8 + 7.7 vs 70.8 + 12.7; p < 0.00). It was concluded that the addition of pentoxifylline to the sperm prepared by the two- layer Percoll gradient method can enhance the sperm membrane integrity, and it may be beneficial to add pentoxifylline to sperm preparation for use in IUI or IVF.

Sperm motility and morphology after preparation using the percoll gradient method compared with the IxaPrep gradient method.

In order to compare the Percell gradient and IxaPrep gradient methods, the percentage of progressively motile sperm and the percentage of sperm with normal morphology were examined. Thirty five normal semen sample were collected from the male partners of infertile couples attending the Infertility Clinic. The samples were divided into two fractions of 1 ml each. Both fractions were processed using the Percoll gradient and IxaPrep gradient methods, and sperm motility and normal morphology were evaluated. The percentages of spermatozoa that showed progressive motility and normal morphology from the sperm preparation using the IxaPrep gradient method were significantly higher than those prepared by the Percoll gradient method (77.7 + 12.0 vs 73.7 + 12.3 and 64.4 + 15.1 vs 61.1 + 13.3 respectively ; p<0.05). It was concluded that sperm preparation using IxaPrep gradient method showed better sperm quality with respect to progressive motility and normal morphology than those using the Percoll gradient method, The IxaPrep gradient method can be considered as an alternative to the Percoll gradient for sperm preparation.

Comparison of human menopausal gonadotrophin and recombinant follicle-stimulating hormone in in-vitro fertilisation and pregnancy outcome.

The study was carried out to compare the effectiveness of human menopausal gonadotrophin (HMG) and recombinant follicle-stimulating hormone (recombinant FSH) in term of in vitro fertilisation (IVF) and pregnancy outcome. A total of 238 patients who underwent IVF for infertility treatment were included in the study. The first attempt of controlled ovarian stimulation was recorded and evaluated. A long protocol of ovarian stimulation was performed with gonadotrophin releasing hormone analogue (GnRH-a) administration. Gonadotrophin, which was either HMG (group A) or recombinant FSH (group B), was administrated to each patient for ovarian stimulation. The results of this study showed no difference in the number of stimulation days, fertilised oocytes, transferred embryos and cycles with embryos available for freezing between the two groups. Although the starting doses of both gonadotrophins were similar, the total dosage of HMG was higher than that of recombinant FSH (48.8ฑ20.8 versus 42.9ฑ20.0, p = 0.03). The number of retrieved oocytes in group A was higher than that in group B (9.5ฑ4.4 versus 8.3ฑ4.3, p = 0.04). The differences in cancellation rate, fertilisation rate, pregnancy rate per cycle and per transfer, as well as implantation rate between the two groups was not statistically significant. In conclusion, patients who underwent ovarian stimulation with GnRH-a long down-regulation still benefit for HMG for their treatment. We did not find any difference in fertilisation rate or pregnancy rate as well as implantation rate between HMG and recombinant FSH. A greater number of oocytes were retrieved in patients treated with HMG. However, more ampoules of HMG were administrated to achieve ovarian stimulation, compared with recombinant FSH.

Comparison of sperm motility between the swim-up and swim-down methods.

The objective of this study was to compare two methods for the extraction of high motility sperm; the swim-up and swim-down techniques. The result showed that both methods produced greater sperm motility than the original semen. The motility of the sperm was lower in the swim-down method compared with the swim-up method (65.5+11.2% vs 76.5+10.1%; p<0.01).