• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.239-244
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• 1998

The purpose of this study is compare IVF cycle outcome in poor responders between clomiphene citrate (CC) stimulated and controlled ovarian hyperstimulation (COH) protocol. A total of 94 patients responding poorly in previous IVF cycles (estradiol<600 pg/ml or less than 3 oocytes retrieved) subsequently underwent either COH (COH group: 122 cycles, 68 patients) or CC-stimulated cycles (CC group: 43 cycles, 26 patients). CC was administered for five consecutive days starting on cycle day 3 at a dose of 100 mg daily. Serial transvaginal ultrasound examination was done from cycle day 8. Urine was collected $3\sim4$ times before hCG injection for the detection of LH surge. The hCG was administered when serum estradiol reached greater than 150 pg/ml and mean follicle diameter>16 mm. In COH group, ovarian stimulation was done using short protocol (GnRH-a/FSH/HMG/hCG). No difference in age or number of transferred embryos was found between CC group and COH group. COH group had significantly (p<0.05) higher mean peak level of $E_2$ ($810{\pm}112$ vs $412{\pm}55$ pg/ml) and greater number of retrieved oocytes ($3.0{\pm}0.2$ vs $2.0{\pm}0.2$) than CC group. CC group had transferred embryos $(1.8{\pm}0.2)$ compared with $(2.1{\pm}0.2)$ in COH group. However, CC group had higher pregnancy rate than COH group per retrieval [26.9% (7/26) vs 6.2% (6/97)], or per transfer [31.8% (7/22) vs 7% (6/86)]. Although cycle cancellation rate in CC group (48.8%) was higher than that of COH group (21.3%), the pregnancy rate per cycle in CC group was still higher (16.3%) than COH group (4.9%). In addition, implantation rate in CC group was 17.5% (7/40), which was significantly (p<0.01) higher than 3.9% (7/180) in COH group. These data suggest that oocyte and embryo quality are lower in COH cycles of poor responders than CC cycles. We suggest that clomiphene citrate stimulated IVF cycle may be more efficient than COH IVF cycle in poor responders in terms of lower costs and higher pregnancy performance.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.29-39
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• 2004

Objectives: This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences. Methods: A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media. Results: A total of 3, 209 oocytes were collected, and 83.8% (2, 212/2, 640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2, 043/2, 071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1, 935/ 2, 043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67). Conclusions: The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.1
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• pp.67-74
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• 2000

Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.