체외수정시술시 난자 세포질내 정자 주입술을 이용한 난자의 미세보조 수정술에 관한 연구

이진용(JY Lee),문신용(SY Moon),김정구(JK Kim),최영민(YM Choi),김석현(SH Kim),오선경(SK Oh),서창석(CS Suh),류범용(BY Ryu) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.6

In spite of much progress in in vitro fertilization and embryo transfer(IVF-ET) program, the pregnancy rate remains at 20~30%, and the endometrial implantation rate per embryo transferred at 10%. Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization(MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after intracytoplasmic sperm injection(ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not be accepted for standard IVF-ET because of extremely impaired semen characteristics(Group A) and because of failure in fertilization of extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles(Group B). From March, 1995 to December, 1996, a total of 114 cycles of IVF-ET with ICSI in 65 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. In Group A, 34 patients were evaluated with semen score such as number of total motile sperms, and then divided into 4 groups accordingly. In 62 ICSI cycles, the number of oocytes retrieved after controlled ovarian hyperstimulation(COH) was 12.4±6.8, and the number of oocytes optimal for ICSI procedure was 8.8±5.5. The fertilization rate of 65.7± 23.6% could be obtained after ICSI. The number of embryos transferred was 4.4±2.2 with the mean CES of 50.5±34.3 in ICSI cycles. The overall pregnancy rate was 24.2%(15/62) per cycle and 44.1%(15/34) per patient. There were no significant differences in the pregnancy rates among 4 groups. Although more mature oocytes were retrieved, the fertilization rate was significantly lower in Group A-1 compared with Group A-IV. However, semen score did not clearly affect the outcomes of ICSI in couples with severe male factor infertility. In Group B, the number of oocytes retrieved after COH was 10.5±6.1 in 49 previous cycles, and 10.8±5.7 in 52 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was 8.5±5.1 with the fertilization rate of 72.4±22.5%. The number of embryos transferred was 1.4±2.4 in previous cycles, and 4.7±1.8 with the mean CES of 50.4 ±29.9 in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 30.8%(16/52) per cycles and 51.6%(16/31) per patients. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with few spermatozoa for the conventional methods of in vitro insemination and with the past history of failure in fertilization or low fertilization rate in the previous cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to the effective management of infertile couples.

미수정 및 저수정율의 기왕력을 지닌 체외수정시술 환자에서의 난자 세포질내 정자 주입술을 이용한 미세보조 수정술에 관한 연구

문신용,김석현,채희동,김광례,이재훈,김희선,류범용,오선경,서창석,최영민,김정구,이진용,Moon, Shin-Yong,Kim, Seok-Hyun,Chae, Hee-Dong,Kim, Kwang-Rye,Lee, Jae-Hoon,Kim, Hee-Sun,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Choi, Young-Min,Kim, 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.1

Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization (MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after partial zona dissection (PZD), subzonal insertion (SUZI), and intracytoplasmic sperm injection (ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not undergo standard IVF-ET repetitively because of failure in fertilization or extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles. From March, 1995 to May, 1996, 27 cycles of IVF-ET with ICSI in 19 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $10.50{\pm}6.13$ in 30 previous cycles, and $10.57{\pm}5.53$ in 27 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was $7.89{\pm}4.30$, and the fertilization rate of $67.9{\pm}20.2%$ could be obtained after ICSI. The number of embryos transferred was $1.43{\pm}2.40$ in previous cycles, and $4.36{\pm}1.77$ with the mean CES of $41.8{\pm}27.4$ in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 29.6% (8/27) per cycle and 42.1% (8/19) per patient with the clinical pregnancy rate of 22.2% (6/27) per cycle and 31.6% (6/19) per patient. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with the past history of failure in fertilization or low fertilization rate of oocytes in the previous IVF-ET cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to effective management of infertile couples.

난자 및 배아성숙이 시험관아기시술 성공률에 미치는 영향

이여일(YI Lee),김재훈(JH Kim),박현정(HJ Park) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.5

There are a number of factors affecting the pregnancy rate in in vitro fertilization (IVF): the etiology of infertility, ages of husband and wife, the type of ovarian stimulation, the number of aspirated oocytes, the maturity of oocyte, embryo quality, the number of fertilized oocytes, the cumulative embryo score, and the number of embryos transferred. Among them the maturity of oocyte and the quality of embryo appear to be the most important factors influencing the pregnancy rate. The aim of this study designed to evaluate the effect of oocyte maturity, embryo quality, the number of embryos transferred, and the cumulative embryo score to fertilization rate on whether the patient conceived or not. A total of 146 consecutive cycles of in vitro fertilization-embryo transfer(IVF-ET) were studied using two ovarian hyperstimulation protocols; follicular stimulating hormone(FSH)/ human menopausal gonadotropin(HMG)/human chorionic gonadotropin(hCG) and gonadotropin releasing hormone agonist(GnRH-a)/FSH/HMG/hCG. Following were the results. 1. The quality and quantity of oocytes and embryos, fertilization rate, and cumulative embryo score between two ovarian hyperstimulation protocols were not different. 2. The more mature oocytes exhibited, the greater fertilization rates in both methods (p<0.01). 3. The average numbers of embryos transferred were 3.65+-0.27 in conceived group and 2.54+-0.18 in nonconceived group, which was statistically higher in conceived group (p<0.001). 4. The pregnancy rate was higher if any matured embryo(Grade I) was present among the embryos transferred(p<0.05). 5. The average cumulative embryo score was 69.70+-6.13 inconceived group and 43.51 +-3.41 in nonconceived group, which was statistically higher in conceived group(p<0.001). These data suggest that the pregnancy rate is higher when the oocyte maturation is Better, the number of embryos transferred is greater, there is at least one matured embryo among the embryos transferred, and the cumulative embryo score is higher.

남성인자 불임환자에서의 체외수정시술시 나자 세포질내 정자 주입술을 이용한 미세보조 수정술의 임상적 효용성에 관한 연구

문신용(SY Moon),김석현(SH Kim),김광례(KR Kim),채희동(HD Chae),이재훈(JH Lee),정경남(KN Chung),김희선(HS Kim),서창석(CS Suh),최영민(YM Choi),김정구(JK Kim),이진용(JY Lee) 대한산부인과학회 1996 Obstetrics & Gynecology Science Vol.39 No.12

The proportion of male factor infertility due to quantitative and qualitative sperm disorders is about 50%~60% in infertile couples. Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many couples with severe andrological problems, and certain couples cannot be even accepted for standard IVF-ET if the number of motile spermatozoa is too low. Recently, several procedures of microassisted fertilization(MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after intracytoplasmic sperm injection(ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF using micromanipulation, and to apply ICSI clinically to infertile couples who could not be accepted for standard IVF-ET because of extremely impaired semen characteristics such as 500,000. In 39 ICSI cycles, the number of oocytes retrieved after controlled ovarian hyperstimulation(COH) was 13.23±6.99, and the number of oocytes optimal for ICSI procedure was 10.07±5.2. The fertilization rate of 59.7±23.1% coould be obtained after ICSI. The number of embryos transferred was 4.26±2.22 with the mean CES of 50.7±30.5 in ICSI cycles. The overall pregnancy rate was 23.1%(9/39) per cycle and 37.5%(9/24) per patient with the clinical pregnancy rate of 15.4%(6/39) per cycle and 25.0%(6/24) per patient. There were no significant differences in the pregnancy rates among 4 groups. Especially, although more mature oocytes were retrieved in Group I compared with Groups III and IV, the fertilization rate and the pregnancy rates were not significantly different. In conclusion, MAF of human oocytes with ICSI could be used successfully for IVF-ET ininfertile couples who had few spermatozoa for the conventional methods of in vitro insemination, and sperm parameters did not clearly affect the outcomes of ICSI in couples with severe male factor infertility. In addition, this development of gamete and embryo micromanipulation will lead to the new potentially exciting techniques such as enucleation for correction of polyspermy, and blastomere biopsy for primplantation genetic diagnosis.

체외수정 배양액에 대한 제대혈청 및 단백질 첨가의 난자성숙 및 난구 세포 분산에 미치는 영향

박현정(HJ Park),신현우(HY Shin),이여일(YI Lee) 대한산부인과학회 1995 Obstetrics & Gynecology Science Vol.38 No.7

OBJECTIVE : To investigate of effect of human cord serum(HCS) and bovine serum albumin(BSA) supplementation on oocyte maturation and cumulus cell expansion in vitro culture media. DESIGN : Random sampling of 6 conceptional media from 7 patients and 8 nonconceptional media from 14 patients resulted from our in vitro fertilization and embryo transfer(IVF-ET) program. Each medium was divided to three groups : Ham`s F-10(unsupplemented), Ham`s F-10 supplemented with 0.4% BSA and Ham`s F-10 supplemented with 10% HCS. Setting, Patients : In vitro fertilization (IVF) program and patients at the IVF clinic of Chonman University Hospital Kwangju. Korea. Interventions : oocyte cumulus cell(OCC) complexes were collected from removed ovaries of pregnant mare serum gonadotropin(PMSG) primed mice, and cultured in three different media. Main outcome Measures : The oocyte maturities were measured by the presence of germinal vesicle breakdown and polar body, and the cumulus cell expansions were analyzed by the degree of cellular dispersion. Results : Oocyte maturity was not significantly different among the three types of media. cumulus cell expansion was significantly increased only by HCS-supplemented media. Comparison of oocyte maturity and cumulus cell expansion among groups showed no significant differences between conceptional and nonconceptional media. Conclusions : The supplementions with HCS or BSA are not able to augment mouse oocyte maturation. The great cumulus expansion in HCS supplemented media suggest the presence of stimulatory in OCC complex. The OCC complex culture provides new insight into qualitative information on prefertilization and perifertilization events.

사정 정자를 이용한 세포질내 정자주입술에서 수정률과 임신율에 영향을 미치는 요인

전진현(JH Jun),임천규(CK Lim),김정욱(JW Kim),변혜경(HK Byun),한미현(MH Han),이호준(HJ Lee),최범채(BC Choi),유근재(KJ Yoo),백은찬(EC Baik),궁미경(MK Koong),손일표(IP Son),강인수(IS Kang),전종영(JY Jun) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.4

Intracytoplasmic sperm injection (ICSI) has been reported as the most successful technique to obtain high fertilization and pregnancy rate in infertile couples who could not be helped by traditional in vitro fertilization or previously described techniques of assisted fertilization. We performed retrospective analysis, in relation to the number of retrieved (NRO) and injected oocytes (NIO), sperm parameters, maturation rate of retrieved oocytes (MRO), fertilization rate after ICSI (FRI) and age of patient (AGP) in 975 ICSI cycles using ejaculated sperm during 1994∼1996. The purpose of this study was to determine the important factors affecting fertilization and ongoing pregnancy rate in the ICSI cycles using ejaculated sperm. A total of 8,809 injected oocytes, 6,138 (69.7%) oocytes were normally fertilized and 213 (22.8%) ongoing pregnancies were achieved in 933 cycles of embryo transfer. The fertilization rates were significantly different in relation to NRO (from 67.3% to 75.0%), NIO (from 68.5% to 73.9%), sperm parameters (from 64.4% to 73.0%), MRO (from 64.6% to 73.9%)and AGP (from 67.8% to 74.4%), respectively. The ongoing pregnancy rates were significantly different in relation to NRO (from 10.0% to 26.6%), NIO (from 10.4% to 27.9%), FRI (from 18.5% to 30.6%) and AGP (from 6.7% to 25.4%), respectively. From these results, it can be concluded that ICSI using ejaculated sperm can provide high normal fertilization rates (above 64%) in all groups analyzed and NRO, NIO, FRI and AGP are important factors determining a successful ongoing pregnancy.

체외수정시술시 반복 실패 불임환자에서 자가면역 항체 측정의 임상적 의의에 관한 연구

김석현(SH Kim),지병철(BC Jee),이희선(HS Lee),서창석(CS Suh),최영민(YM Choi),김정구(JK Kim),문신용(SY Moon),이진용(JY Lee) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.6

The purpose of this study was to investigate the prevalence of serum autoimmune antibodies in patients undergoing in vitro fertilization and embryo transfer (IVF-ET), and to clarify serum autoantibodies as a possible cause of repeated failure in IVF-ET. Sera from a total of 45 infertile patients, who have just enlisted and/or undergone IVF-ET previously, were assayed to identify autoantibodies as followings: lupus anti-coagulant (LAC), anti-double stranded DNA antibody (anti-dsDNA Ab), antinuclear antibody (ANA), rheumatoid factor (RF), antithyroglobulin antibody (anti-TG Ab), and antimicrosomal antibody (anti-MS Ab). The overall prevalence rate of serum autoantibodies was 40.0% (18/45) in 45 IVF-ET patients with the highest rate of 22.2% (10/45) in ANA. The positive rate was similar without statistical significance in both tubal and unexplained infertility groups (32.0% vs. 57.1%). There was a tendency of decreasing prevalence rate with the increasing number of failed cycles in IVF-ET: Group of repeated IVF-ET failure (3 or more cycles) showed significantly lower prevalence rate than Group of 1∼2 failed cycles (15.4% vs. 90.0%, p<0.01). There were no significant differences in age of patients, duration of infertility, number of previously failed cycles of IVF-ET, previous history of conception and viable pregnancy, infertility factor, and clinical pregnancy over 20 weeks between positive and negative serum autoantibodies groups. In conclusion, the presence of serum autoantibodies does not mean a poorer prognosis in IVF-ET, and the prevalence rate of serum autoantibodies in repeated IVF-ET failure patients may just reflect that in general infertile patients.

체외수정시술시 배아의 보조부화술을 이용한 임신율 향상에 관한 연구

김석현,김광례,채희동,이재훈,김희선,류범용,오선경,서창석,최영민,김정구,문신용,이진용,Kim, Seok-Hyun,Kim, Kwang-Rye,Chae, Hee-Dong,Lee, Jae-Hoon,Kim, Hee-Sun,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Choi, Young-Min,Kim, Jung-Gu,Moon, S 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.1

In spite of much progress in vitro fertilization and embryo transfer (IVF-ET) program, the pregnancy rate remains at 20-30%, and the endometrial implantation rate per embryo transferred at 10-15%. As a result, about 90% of embryos may fail to implant to the endometrium, and many attempts such as optimization of follicular development, improvement of in vitro culture system including coculture, and micromanipulation of zona pellucida have been made to improve embryonic implantation after IVF-ET. Recently, several procedures of assisted hatching (AH) using micromanipulation have been introduced, and pregnancies and births have been obtained after AH. To develop and establish AH as an effective procedure to improve embryonic implantation, AH with partial zona dissection (PZD) was performed in 116 cycles of 89 infertile couples who had previous repeated failures of standard IVF-ET more than two times (Group I: 71 cycles in 54 patients), or who had implantation failure of embryos with good quality (Group II: 15 cycles in 13), or who had undergone AH without specific indication (Group III: 30 cycles in 22) from January, 1995 to Februry, 1996, and the outcomes of AH were analyzed according to pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $9.9{\pm}7.1$ in Group I, $11.5{\pm}4.5$ in Group II, and $7.9{\pm}6.4$ in Group III. The number of embryos transferred after AH was $4.7{\pm}1.8$ in Group I, $5.3{\pm}1.3$ in Group II, and $3.5{\pm}2.4$ in Group III. The mean cumulative embryo score (CES) was $56.8{\pm}30.0$ in Group I, $76.1{\pm}35.9$ in Group II, and $38.5{\pm}29.9$ in Group III. The overall clinical pregnancy rate per cycle and per patient was 12.7% (9/71) and 16.7% (9/54) in Group I, 33.3% (5/15) and 38.5% (5/13) in Group II, and 6.7% (2/30) and 9.1% (2/22) in Group III, respectively. There were significant differences in the numbers of oocytes retrieved and embryos transferred, CES, and the clinical pregnancy rate per cycle among three groups. There was a significant inverse correlation between basal serum FSH level and CES, and no pregnancy occurred in patients with CES less than 20. In conclusion, AH of human embryos with PZD prior to ET has improved the implantation and pregnancy rates in IVF-ET patients with the past history of repeated failures, especially in spite of transfer of embryos with good quality, and AH will provide a range of novel techniques which may contribute much to effective management of infertile couples.

체외수정시술을 위한 과배란유도시 Follicle-Stimulating Hormone과 Human Menopausal Gonadotropin의 효용성 비교 연구

김석현(SH Kim),지병철(BC Jee),노재숙(JS Roh),이재훈(JH Lee),서창석(CS Suh),최영민(YM Choi),신창재(CJ Shin),김정구(JK Kim),문신용(SY Moon),이진용(JY Lee),장윤석(YS Chang) 대한산부인과학회 1997 Obstetrics & Gynecology Science Vol.40 No.1

Recent evidence suggests that a high level of serum LH during follicular recruitment and development is associated with poor reproductive outcome. Consequently, exogenous LH administration for controlled ovarian hyperstimulation(COH) in in vitro fertilization and embryo transfer(IVF-ET) may be harmful to folliculogenesis. The purpose of this clinical study was to evaluate and compare the efficacy of human menopausal gonadotropin(hMG) and human follicle-stimulating hormone(hFSH) for COH with long protocol of gonadotropin-releasing hormone(GnRH) agonist in IVF-ET program. Randomized clinical trial was performed in 125 patiens undergoing IVF-ET at Seoul National University Hospital from May to Septebmer, 1995. The inclusion criteria of patients included age<40 years and normal semen analysis, and the study population was also classified into two groups by the etiology of infertility : Group T - 95 patients with only tubal factor and Group O - 30 patients with endometriosis or anovulatory factor. There were no statistically significant differences in dosage(29.0±7.9 vs 26.0±6.8 ampoules) and duration(12.3±1.3 vs 12.2±1.5 days) of gonadotropin administration, serum E2 level on hCG day(1,943±1,255 vs 1,580±1,067 pg/mL), cancellation rate(7.5% vs 6.7%), number of oocytes retrieved(9.9±6.0 vs 11.3±6.0), fertilization rate(68.4% vs 64.5%), number of embryos transferred(4.7±2.0 vs 4.7±2.0), and preganancy rate per cycle(26.3% vs 24.4%) and per ET(28.4% vs 26.2%) between hMG(N=80) and hFSH(N=45) Groups. In Group T, no significant differnces in results of IVF-ET were also detected between hMG (N=61) and hFSH(N=34) Groups. In Group O, serum E2 level on hCG day was significantly higher in hMG Group (N=19) compared with hFSH Group(N=11), but other results of IVF-ET were similar in both Groups. As this study could not demonstrate any significant differences in results of IVF-ET between hMG and hFSH when used for COH in IVF-ET program, it could be concluded that hFSH is at least as efficacious as hMG for COH.

난자 세포질내 정자 주입술을 이용한 체외수정시술시 누적임신율에 관한 연구

김석현(Seok Hyun Kim),심순섭(Soon Sup Shim),지병철(Byung Chul Jee),최성미(Sung Mi Choi),김희선(Hee Sun Kim),류범용(Buom Yong Ryu),오선경(Sun Kyung Oh),서창석(Chang Suk Suh),최영민(Young Min Choi),배광범(Kwang Bum Bai),김정구(Jung Gu 대한산부인과학회 2001 Obstetrics & Gynecology Science Vol.44 No.3

Objective : To evaluate the cumulative pregnancy rate(CPR) of in vitro fertilization and embryo transfer(IVF-ET) with intracytoplasmic sperm injection(ICSI). Methods : Medical records of 260 infertile patients undergoing 519 cycles of IVF-ET with ICSI from January, 1994 to December, 1999 were retrospectively reviewed. The CPR beyond 12 weeks of gestation was estimated by Kaplan-Meier method. The CPRs were compared by log-rank test between groups divided by age of patients, indication of ICSI, and method of sperm retrieval for ICSI. Results : As 70 patients achieved an on-going pregnancy after IVF-ET with ICSI, the PR was 26.9% per patient and 13.5% per cycle. The overall CPR was 54.9% after 6 cycles of IVF-ET with ICSI. As expected, age had a significant strong effect on the CPR; CPRs afer 4 cycles of ICSI were 61.8% in the age group of 30 years(n=81), 43.7% in 31-35 years(n=106), and 15.3% in 36 years(n=73). There was no significant difference in the CPR between abnormal semen analysis group(n=184) and prior low fertilization rate group(n=66). In abnormal semen analysis group, the CPR of surgically retrieved sperm subgroup(n=60) was not significantly different from that of ejaculated sperm subgroup(n=124). Conclusions : The CPR of IVF-ET with ICSI was presented, and it could be of much help in the clinical counseling of IVF-ET patients. ICSI technique could be used successfully for IVF-ET in infertile couples who had the male factor infertility or the past history of low fertilization rate in the previous cycles.