Mouse blastocyst vitrification compared with the conventional slow-freezing method.

To compare mouse blastocyst survival after cryopreservation with vitrification and the slow-freezing method, one-hundred and forty-eight in vitro mouse blastocysts were randomly frozen by the two methods: vitrification and conventional slow-freezing. After being thawed, the blastocysts were assessed for survival and hatching rate. The survival rates of blastocysts cryopreserved by vitrification and slow-freezing were 68.33 and 65.52 per cent (p = 0.89), whereas hatching rates were 51.22 and 44.74 per cent, respectively (p = 0.64). Therefore, vitrification of blastocyst-stage-embryos may be a useful, economic method for freezing the excess blastocysts in some centers where blastocysts are routinely transferred.

The regulation of assisted reproductive technology in Thailand.

The Executive Board of the Medical Council of Thailand has set up an ad hoc committee to establish the regulations of practising of assisted reproductive technology. The committee assigned the Royal Thai College of Obstetricians and Gynecologists to supervise and take charge of the administration and formulate an annual report in accordance with the Thai Medical Council Declaration. The regulation was finally approved on October 9, 1997. It was announced in the Royal Gazette on December 26, 1997 and since then the prescription of standard measures for ART practice has been effected.

Correlation between human follicular diameter and oocyte recovery, metaphase II oocytes and fertilization rate in intracytoplasmic sperm injection programs.

This prospective study was undertaken to establish the correlation between the follicular diameter, oocyte recovery, metaphase II (MII) oocyte, and fertilization rate in intracytoplasmic sperm injection (ICSI) programs. Thirty one patients undergoing 31 ICSI cycles from August 1998 to January 1999 at King Chulalongkorn Memorial Hospital were studied. The patients were stimulated with either short or long GnRHa portocol. During transvaginal aspiration of follicles, follicular diameters were measured. The maturation stage of oocytes and fertilization rate were evaluated. Four hundred and seventy eight follicles were measured and classified into 3 groups, group A (<10 mm), group B (10-14 mm) and group C (>14 mm). The oocyte recovery rate and MII oocytes were different between groups A, B, and C but the fertilization rate was not different. In conclusion, the follicular diameter correlated with oocyte recovery and oocyte maturation but not with the fertilization rate in ICSI programs. The quality of embryos was lower in oocytes from small follicles.

Effects of contaminated endometriotic contents on quality of oocytes.

The mechanism of infertility associated with endometriosis is poorly understood. There is evidence supporting that women with ovarian endometriosis have a lower pregnancy rate than women with peritoneal lesions only. This study aimed to evaluate the effects of endometriotic contents contamination while retrieving oocytes on oocytes' quality. Thirty-eight infertile patients with endometriotic cysts from January 1993 to June 2000 were enrolled in this study. There were no statistically significant differences among the quality of oocytes and embryos from the contaminated, non-contaminated, and control group. However, the fertilization rate and pregnancy rate were impaired by the contamination of endometriotic contents. We conclude that ovarian endometriosis should be treated before starting in vitro fertilization program in order to increase the pregnancy outcome.

Effects of different biopsy methods on the development of preimplantation mouse embryos, in vitro and in vivo: implication for preimplantation genetic diagnosis.

The aim of this study was to investigate the efficacy of two different embryo biopsy techniques, direct aspiration and partial zona dissection (PZD)-push, on subsequent in vitro and in vivo development of 8-cell stage mouse embryos. It was found that the rates of normal blastocyst formation and hatching blastocysts of direct aspiration, PZD-push, solution control and control embryos were not significantly different (80.8%, 81.6%, 84.5%, 86.7% and 71.9%, 72.3% and 74.6%) respectively. There was, however, a significant reduction in rate of complete hatching blastocysts (P < 0.1) (72.9% aspiration versus 85.2 per cent solution control and 86.4% control) and rate of live-born fetuses (24.2% aspiration versus 43.3% solution control and 41.2% control) (P < 0.05) in the direct aspiration group but no significant difference in the PZD-push group (80.3% of complete hatched blastocysts and 33.8% of live-born fetuses). These findings indicated that embryo biopsy with PZD-push was superior to the direct aspiration method. This mouse embryo biopsy model was useful in advancing development of biopsy technique for human preimplantation genetic diagnosis.

Clinical effects of gestrinone for the treatment of pelvic endometriosis in infertile patients.

The clinical efficacy and side effects of oral gestrinone, 2.5 mg twice weekly, were evaluated in this prospective study involving 22 patients with laparoscopically confirmed endometriosis. All patients came to the hospital with infertility problem. After 6 months of treatment, 81 per cent of patients had amenorrhea. Dysmenorrhea and pelvic pain were reduced from 90.9 per cent to 14.3 per cent and 81.8 per cent to 9.5 per cent respectively. Pelvic tenderness and induration were improved from 55.6 per cent and 50.0 per cent to 15.8 per cent and 10.5 per cent respectively. Androgenic effects such as acne was founded in 18.2 per cent of the patients. Return of fertility was observed in 25 per cent (5 patients) after 30-254 days post treatment. No serious side effect was detected during the treatment. The results suggest that gestrinone may be considered an option for the treatment of endometriosis related infertility.

Successful pregnancy in a case of azoospermia from congenital bilateral absence of vas deferens (CBAVD) using percutaneous epididymal sperm aspiration (PESA) and intracytoplasmic sperm injection (ICSI).

In men with obstructive azoospermia, bypass surgery would obviously be the most acceptable form of treatment as it gives the couple an opportunity to conceive naturally. However, when this has failed to restore patency or when surgery is not feasible (congenital absence of vas), fertility treatment using spermatozoa aspirated from the epididymis should be considered. Percutaneous epididymal sperm aspiration (PESA) is more acceptable to patients than micro epididymal sperm aspiration (MESA) because it eliminates the requirement for a general anaesthetic, post-operative pain, and the risk of haematoma formation, thus allowing a rapid return to normal activity of the husband. To our knowledge, this is the first reported case in Thailand where a pregnancy resulted in a couple whose infertility was due to azoospermia from the congenital absence of vas deferens. Spermatozoa collected through PESA were used in ICSI to achieve fertilization. The PESA technique, due to its simplicity is the choice of treatment for obstructive azoospermia.

Reliable sex determination of mouse preimplantation embryos by PCR amplification of male-specific genes in single blastomeres.

OBJECTIVE: To assess the reliability of sex determination in mouse preimplantation embryos using the two-step polymerase chain reaction method. SETTING: Division of Immunology, Department of Microbiology and Division of Reproductive Medicine, Department of OB/GYN. METHODS: The Sry and Zfy genes, known to be present in the sex-determining region of mouse Y chromosome, were selected for Y-specific target sequences and DXNds 3 locus located on mouse X chromosome was served as the internal control sequence. DNAs extracted from heart blood of male and female mice were used to test the correctness and specificity of the selected primers using the two-step PCR method. The same experimental conditions were then used to amplify the single copy genes in single mouse blastomeres with two pairs of primers for each of the target sequences. The sex-determined embryos were transferred to the uteri of pseudopregnant recipients to test the consistency of the assay system. RESULTS: All male and female blood DNA sample results confirmed the correct sex identification of the origin (100%). Nineteen of 20 single blastomeres showed the accurate diagnosis when compared with theirs 7/8 embryos. The sex of 36 of 37 mouse pups born from biopsied male and female embryos agreed with the predicted sex. CONCLUSION: The reliable genetic analysis of sex chromosome- specific sequences in single cell is possible by the two-step PCR method and could be applied for diagnosis of defective genes of human preimplantation embryos derived from the in vitro fertilization program.

Basal follicle--stimulating hormone levels on day 3 of previous cycle are predictive of in vitro fertilization outcome.

Serum FSH level may be an indicator of the functional potential of the ovary. The basic control of LH and FSH is by a negative feedback system involving the hypothalamus, anterior pituitary, and ovary. Basal serum FSH on day 3 of previous cycles in 36 patients underwent ovarian stimulation with a combination of GnRH agonist and FSH and hMG in an IVF program were evaluated. The results in this study show that a high basal serum FSH level is associated with a higher cancellation rate, a lower peak serum E2 level, lower number of oocytes retrieved and a lower clinical pregnancy rate.