자궁내막증과 동반된 불임환자의 보조생식술을 이용한 치료

문신용,이경순,노재숙,서창석,김석현,최영민,신창재,김정구,이진용,장윤석,Moon, Shin-Yong,Lee, Kyung-Soon,Roh, Jae-Sook,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Young,Chang, Yoon-Seok 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

Though the endometriosis is not always related with infertility, endometriosis causes infertility in some patients. There are many treatment modalities of infertile patients who have endometriosis. In recent years, Assisted Reproductive Technology(ART) have been widely accepted as being a useful tool for the treatment of infertile endometriotic patients. The objective of this study was to evaluate the outcome of ART in infertile endometriotic patients who have been carried out IVF-ET from Jan, 1992 to Dec, 1994 and to compare the results between COH/IUI and IVF-ET in the patients with endometriosis stage I. Tubal disease only patients were grouped(308 patient, 956 cycles) as a control. Endometriosis group was subdivided into 4 groups according to American Fertility Society classification; endometriosis stage I (45 patients, 61 cycles), stage II (26 patients, 39 cycles), stage III (26 pateitns, 37 cycles), stage IV (33 patients, 50 cycles). The outcomes of IVF-ET in endometriosis patients were as follows; The oocyte recovery rates were significantly lower in stage III, IV endometriosis. In case of stage III endometriosis, the fertilization rate was significantly lower than other stages of endometriosis. Clinical pregnancy rates per cycle were not different between the tubal group(22%) and the endometriosis group(25%). According to endometriosis stage, the implantation rate and clinical pregnancy rate were significantly lower in stage IV (5.6%, 16%) compared with other stages (I; 10.0%, 26%, II;9.8%, 31%, III;12.6%, 32%). It suggests that some factor like autoantibodies may inhibit implantation of embryos in stage IV endometriosis. To evaluate the possibility that simply increasing the number of gametes at the site of fertilization might account for pregnancies attributed to IVF-ET, the authors retrospectively analyzed the outcome of couples undergoing IUI during hMG cycles and CC cycles between 1992 and 1994 in the women with endometriosis stage 1. In case of stage I endometriosis, though the COH/IUI group showed lower FSH level and lesser age profile than IVF-ET group, IUI group has resulted in lower pregnancy rates(19.2%) compared with the IVF-ET group(26.2%). In conclusion, endometriotic infertile patients can get comparable pregnancy rates with the tubal factor infertility patients during IVF-ET program. Moreover even in stage I endometriosis, IVF-ET may be an more effective treatment modality than COH/IUI.

체외수정시술을 위한 과배란유도에 있어 GnRH Antagonist의 임상적 효용성과 혈중 호르몬 농도의 변화

문신용,천은경,김상돈,최영식,지병철,구승엽,서창석,최영민,김정구,김석현,Moon, Shin-Yong,Chun, Eun-Kyung,Kim, Sang-Don,Choi, Young-Sik,Jee, Byung-Chul,Ku, Seung-Yup,Suh, Chang-Suk,Choi, Young-Min,Kim, Jung-Gu,Kim, Seok-Hyun 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.4

Objectives: To evaluate the efficacy of GnRH antagonist cetrorelix in women undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) and to determine changes in serum hormone concentrations during cetrorelix administration. Methods: We performed a clinical trial on 30 patients undergoing COH with highly purified follicular stimulating hormone (HP-FSH) and gonadotropin releasing hormone antagonist (GnRHant), cetrorelix. FSH was administrated from day 2 or 3 of cycle with fixed dose and adjusted according to individual response. 0.25 mg of cetrorelix was injected daily subcutaneously from stimulation day 5 until the day of hCG administration. Daily ultrasound monitoring was performed for growing follicles and serum levels of luteinizing hormone (LH), estradiol ($E_2$) and progesterone were measured daily during cetrorelix administration. Up to 4 embryos were transferred. Results: Mean age of enrolled patients was $32.0{\pm}3.4$ years (mean $\pm$ S.D.). All of 30 patients underwent oocyte pick-up, and embryo transfer was done in 28 patients. The total and mean numbers of received oocytes were 196 and $6.5{\pm}4.7$, the number of fertilized eggs was 111, and the fertilization rate was 56.6%. Total duration of FSH administration was $9.2{\pm}2.2$ days and mean of $24.3{\pm}7.7$ ampules of HP-FSH was administered. Total duration of cetrorelix administration was $5.7{\pm}1.9$ days. Serum LH and progesterone levels were maintained in the range of $1.4{\sim}2.9\;mIU/mL$ and $0.3{\sim}0.6\;ng/mL$, which respectively reflected effective prevention of premature LH surge. Clinical pregnancies were achieved in 9 patients, and overall clinical pregnancy rate was 30.0% per oocyte retrieval, and 32.1% per embryo transfer. Conclusion: GnRH antagonist is safe and convenient for COH for IVF-ET and effective with optimal pregnancy rate.

Klinefelter 증후군 및 Y염색체 이상으로 진단된 환자들의 염색체 결과 및 임상 양상

문신용,최석태,고희정,오선경,한정호,서창석,김석현,최영민,김정구,이진용,Moon, Shin-Yong,Choi, Seok-Tae,Ko, Hee-Jung,Oh, Sun-Kyung,Han, Jung-Ho,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Lee, Jin-Yong 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

Klinefelter's syndrome is a very important disease in gynecologic endocrinologic fields, because the patients with this karyotype complain of infertility, azospermia and ambiguous genitalia. Y chromosome is an important chromosome which determine genetic sex and the structure of gonad and genitalia. In this study, to elucidate the cytogenetic characteristics and clinical features of Klinefelter's syndrome and Y chromosomal abnormalities in Korea, we studied 303 cases of Klinefelter's syndrome and 11 cases of Y chromosomal abnormalities which were diagnosed by chromosomal analysis at the Cytogenetic Laboratory, Institute of Reproductive Medicine and Population, Seoul National University for 12 years from January 1984 to December 1996. The results of this study showed as follows: 1. In a total of 9275 cases, there were 303 cases (3.3%) of Klinefelter's syndromes, 11 cases (0.1%) of Y chromosomal abnormalities. 2. In 102 cases of patients showed typical clinical features of Klinefelter's syndrome, 101 cases (99%) of them were diagnosed to Klinefelter's syndrome in karyotyping. 3. In 303 cases of Klinefelter's syndrome, there were 277 cases (91.4%) of 47,XXY complement, 16 cases (5.3%) of mosaicism, 2 cases (0.7%) of 48,XXXY, 5 cases (1.7%) of 48,XXYY and 3 cases (1.0%) of 49,XXXXY. 4. In 303 cases of Klinefelter's syndrome, 284 cases (93.7%) of them were diagnosed after puberty and only 19 cases (6.3%) of them were diagnosed before puberty. 5. In 303 cases of Klinefelter's syndrome, there were 146 cases (48.2%) of patients with infertility-associated chief complaints, 101 cases (33.3%) of patients with typical clinical features of Klinefelter's syndrome, 22 cases (7.3%) of patients with ambiguous genitalia. 6. In patients with Klinefelter's syndrome, 48,XXYY and 49,XXXXY had serious symptoms such as mental retardation, developmental delay, Down syndrome-like features, congenital anomalies, but 48,XXYY and other mosaicisms had infertility-associated symptoms or ambiguous genitalia. 7. The 8 cases of polysomy Y (XYY complement) showed several serious symptoms such as Down syndrome-like features, mental retardation, fragile X syndrome-like feature, congenital anomalies, ambiguous genitalia which could be detected before puberty.

인간 정자의 생식력 평가에 있어 첨제반응율과 햄스터 난자 침투 분석법의 비교연구

문신용,류범용,오선경,서창석,김석현,최영민,신창재,김정구,장윤석,이진용,Moon, Shin-Yong,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Chang, Yoon-Seok,Lee, Jin-Yong 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

This study was designed to determine the relationship between sperm acrosome reaction following ionophore challenge(ARIC) and hamster ovum sperm penetration assay(SPA) as assessment of fertilizing capacity of male. ARIC test and SPA were performed in 23 fertile and 19 subfertile men. The results were as follows; Sperm concentration was significantly higher in fertile group compared with subfertile group: $114.6{\pm}64.40$ vs $61.3{\pm}46.50{\times}10^6/ml$. However, there were no significantly differences in seminal volume, motility and motility index, respectively. There was a significantly correlation between spontaneous and induced AR in fertile and subfertile group, respectively. ARIC value was significantly higher in fertile group, compared with subfertile group: $12.0{\pm}5.57%$ vs $2.6{\pm}4.96%$. Both Penetration rate(PR) and Penetration index(PI) were significantly higher in fertile group, compared with subfertile group: $97.4{\pm}7.40%$ vs $64.9{\pm}36$. 20% and $5.4{\pm}2.88$ vs $1.5{\pm}1.47$, respectively. The Positive predictive value(PPV), Negative predictive value(NPV), sensitivity and specificity of ARIC test (cut-off: 8.5) and SPA(PI cut-off : 3.0) in predicting fertility were 95.0%, 81.8%, 82.6%, 94.7% and 95.2%, 85.7%, 87.0% and 94.7%, respectively. There was no significantly difference in predicting fertility between ARIC test and SPA. In conclusion, ARIC test was shown to have a predictive value for fertilizing capacity comparable to that of the hamster ovum sperm penetration assay. Therefore, ARIC test may be a simple and cost-effective addition to existing semenology instead of SPA.

체외수정 생쥐 배아에서의 배아 줄기세포 확립

문신용,박용빈,김희선,오선경,천대우,서창석,최영민,김정구,이진용,김석현,Shin, Yong-Moon,Park, Yong-Bin,Kim, Hee-Sun,Oh, Sun-Kyung,Chun, Dae-Woo,Suh, Chang-Suk,Choe, Young-Min,Kim, Jung-Gu,Lee, Jin-Yong,Kim, Seok-Hyun 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.1

Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

인간 배아의 동결보존에 관한 연구

문신용,김정훈,김석현,최영민,신창재,김정구,이진용,장윤석,Moon, Shin-Yong,Kim, Chung-Hoon,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.2

Controlled ovarian hyperstimulation(COH) for in vitro fertilization and embryo transfer(IVFET) often results in the production of more embryos than can be efficaciously transferred at one time. However, embryo cryopreservation provides a mechanism by which additional embryos can be stored for later thawing and transfer. From November, 1990 to October, 1992, we completed 42 transfer cycles of cryopreserved pronucleus(PN) l-cell embryos using the fixed protocol of hormonal replacement therapy in a physiological manner regardless of individual ovarian function. Artificial endometrial stimulation was performed with only exogenous estradiol and progesterone(E-P) in 36 transfer cycles (Group I) and with gonadotropin-releasing hormone agonist(GnRHa) and exogenous estradiol and progesterone(GEEP) in 6 transfer cycles(Group II ). The results were as follows. 1. The Survival rate of total cryopreserved-thawed embryos was 64.9%(198/305): 64.9% (172/265) in Group I and 65.0% (26/40) in Group II. 2. Total 168 embryos were transferred with an average of 4.7 per ET in Group I and total 26 embryos were transferred with an average of 4.3 per ET in Group II. 3. The pregnancy rate(PR) per cryopreserved-thawed ET and the implantation rate was 33.3 %(14/42) and 6.7%(13/194), respectively. The PRs per cryopreserved-thawed ET were 30.6% (11/36) in Group I and 50.0% (3/6) in Group II without significant difference. 4. The take home baby rate was 11.1%(4/36) in Group I and 33.3% (2/6) in Group II.

과배란유도시 난포기 LH의 생물학적 활성이 성숙난자의 수정율 및 난할율에 미치는 영향에 관한 연구

문신용,김석현,황태영,신창재,이진용,장윤석,Moon, Shin-Yong,Kim, Seok-Hyun,Hwang, Tae-Young,Shin, Chang-Jae,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.1

To investigate the factors that affect the fertilization and cleavage rates of mature oocytes, 44 patients undergoing controlled ovarian hyperstimulation(COH) with FSH/hMG/hCG regimen for IVF - ET were analyzed. During follicular phase, serum LH levels were measured by radioimmunoassay and bioassay. Based on the mean follicular immunoactive LH(i-LH) and bioactive LH(b-LH) levels, patients were divided into 3 groups, respectively. There were no significant differences in basal serum FSH levels on menstrual cycle day 3, serum estradiol($E_2$) and progesterone ($P_4$) levels on the day of hCG administration, and the numbers of follicles aspirated and oocytes retrieved among groups. In relation to the mean follicular i-LH levels, the fertilization and cleavage rates of mature oocytes did not show a significant difference among groups. However, in groups with higher mean follicular b-LH levels, the fertilization and cleavage rates were reduced significantly. During late follicular phase, day-to-day variance in b-LH levels was not significant, but there was a significant difference among groups. There was no significant correlation between serum P. and b-LH levels. These data suggest that the fertilization and cleavage rates of mature oocytes are adversely affected by the raised mean follicular b-LH levels, and monitoring of serum b-LH levels is more useful in COH when compared with i-LH. It appears that the reduced rates are not due to the attenuated endogenous LH surge.

생쥐배아의 부화와 탈각에 미치는 Pronase의 영향

문신용,최성미,김희선,류범용,오선경,서창석,김석현,최영민,김정구,최규홍,이진용,Moon, Shin-Yong,Choi, Sung-Mi,Kim, Hee-Sun,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Choi, Kyu-Hong,Lee, Jin-Y 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.

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.

Clomiphene Citrate 부하검사와 난소 반응 예측 인자와의 연관성에 관한 연구

문신용,채희동,김광례,서창석,김석현,최영민,신창재,김정구,이진용,Moon, Shin-Yong,Chae, Hee-Dong,Kim, Kwang-Rye,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Yong 대한생식의학회 1996 Clinical and Experimental Reproductive Medicine Vol.23 No.3

Objective: To determine the cutoff value of clomiphene citrate challenge test(CCCT) that can predict the normal and abnormal(diminished) ovarian response group and to assess the usefulness of CCCT as a predictor of ovarian reserve. Materials and Methods: From March 1994 to Februry 1996, CCCT was performed to 129 infertile patients and among them, 20 patients whose basal FSH on the third day of menstrual cycle was more than 20 mIU/ml were excluded. At the same time, the same CCCT was performed to the fifteen healthy volunteers with proven fertility to determine the cutoff value of CCCT. Results; 1) A FSH value higher than 23.4 mIU/ml, measured on the 10th day of menstrual cycle, was defined as a abnormal ovarian response. The cutoff value of 23.4 mIU/ml is more than 2 standard deviations(SD) above the mean value of 15 healthy women after CCCT. 2) The abnormal CCCT group, the subpopulation with a FSH value of 23.4 mIU/ml or more, was 7.3%(8/109) and their mean age was higher than the normal CCCT group($36.5{\pm}4.5$ vs. $32.9{\pm}4.8$, P = 0.059). And the percentage of the patients older than 35 years of the abnormal CCCT group was significantly higher than that of the normal CCCT group(62.5% vs. 38.6%, p <0.05). 3) There was no correlation between the hormone values of the third day and the 10th day of menstrual cycle before and after CCCT except between FSH of the third day and the 10th day. Conclusion: The CCCT is a good method to predict the individual ovarian response to COH for ART, especially in the patients who has no other abnormal findings that predict poor prognosis. And it is neccessary to determine the cutoff value of CCCT by the large numbers of randomized study, and combining the previously proven prognostic factors, it can be applicated in many individual centers for evaluate the ovarian response before ART program.