다운증후군의 세포유전학적 연구

문신용,박중신,최진,고희정,오선경,신창재,Moon, S.Y.,Park, J.S.,Choe, J.,Ko, H.J.,Oh, S.K.,Shin, C.J. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.2

A study has been carried out to elucidate the cytogenetic characteristics of Down's syndrome in Korea. This study includes 877 cases which were diagnosed as Down's syndrome by the chromosomal analyses at the Cytogenetic Laboratory, Institute of Reproductive Medicine and Population, Seoul National University for 13 years from January, 1984 to December, 1996. 1. 83.6% of cases were diagnosed under 1 year of age and 10.9% were between 1 and 4 years old. The overall sex ratio was 3 to 2 (male to female). 2. The most frequent indication for cytogenetic analyses was suspicion of Down's syndrome. The next were growth retardation, congenital heart diseases, congenital anomalies. 3. 88.4% of cases had free trisomy 21. In 6.5%, there was translocation, mostly Robertsonian t(14;21) or t(21;21). 3.9% of cases were mosaics mostly with one normal cell line. 4. Karyotyping was also performed in 204 parents of patients. 6 parents (2.9%) were seen to be translocation carriers of Down's syndrome. We find the unique features of Down's syndrome in Korea that the incidences of free trisomy 21 is relatively lower and that translocation is higher than western countries.

동결보존된 부고환 정자로 ICSI 시술 후 수정된 수정란의 동결보전 및 배아이식에 의한 임신 1례

문신용,이희선,김희선,류범용,방명걸,오선경,서창석,김석현,최영민,김정구,이진용,Moon, S.Y.,Lee, H.S.,Kim, H.S.,Ryu, B.Y.,Pang, M.G.,Oh, S.K.,Suh, C.S.,Kim, S.H.,Choi, Y.M.,Kim, J.G.,Lee, J.Y. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.2

This case report describes the pregnancy following the transfer of cryopreserved embryos generated from intracytoplasmic sperm injection (ICSI) using frozen-thawed sperm obtained by microepididymal sperm aspiration (MESA) in patient with congenital absence of the vas deferens (CAVD).

난소 난종 환자에서의 Estrogen-Progesterone 치료 및 질식 난소 낭종 천자에 관한 연구

문신용,김석현,황태영,신창재,김정구,이진용,장윤석,Moon, S.Y.,Kim, S.H.,Hwang, T.Y.,Shin, C.J.,Kim, J.G.,Lee, J.Y.,Chang, Y.S. 대한생식의학회 1989 Clinical and Experimental Reproductive Medicine Vol.16 No.1

Some infertile patients who need IVF-ET for conception have small ovarian cysts diagnosed in pelvic ultrasonography. It is well known that it is impossible or very difficult to perform controlled ovarian hyperstimulation(COH) for such patients because of the poor ovarian response or the possibility of ovarian hyperstimulation syndrome(OHSS). To remove or to decrease the size of ovarian cysts, estrogen and progesterone (E-P) therapy with oral contraceptives for 2 cycles and transvaginal aspiration of ovarian cysts using transvaginal ultrasonography were performed in 36 IVF-cancelled infertile patients with ovarian cysts from February to October, 1988 at Seoul National University Hospital. Thirty-nine ovarian cysts($32.8{\pm}9.6$mm in mean diameter) were treated with E-P therapy, and their size decreased to $28.2{\pm}11.0mm$ after 1 cycle and significantly to $24.8{\pm}14.7mm$ after 2 cycles. After E-P therapy for 2 cycles, 7(17.9%) ovarian cysts disappeared in ultrasonography, 9(23.1%) decreased in size significantly, 18(46.2%) had no change in size and 5(12.8%) increased in size. Thirty-two ovarian cysts($30.2{\pm}9.7mm$) in 30 patients were aspirated transvaginally, and there was no significant decrease in size after follow-up transvaginal ultrasonography($27.8{\pm}12.5mm$). After transvaginal aspiration, 3(9.4%) ovarian cysts disappeared and 28(87.5%) had no change in size. The mean amount of the transvaginally aspirated cystic fluids was $19.6{\pm}13.2ml$, and there was no malignant cells in aspiration cytology. Four endometrioid cysts, one dermoid cyst and one mucinous cyst could be diagnosed in consideration of the findings of transvaginal ultrasonography and the characteristics and cytology of aspirated fluids. Therefore E-P therapy and transvaginal aspiration of ovarian cysts had made it possible to restart IVF program earlier in the IVF-cancelled patients with ovarian cysts.

생쥐 2-세포배아에 의한 시험관아기 배양용 대아제대혈청의 절적평가에 관한 연구

문신용,신창재,정구민,오선경,방명걸,장윤석,Moon, S.Y.,Shin, C.J.,Chung, K.M.,Oh, S.K.,Pang, M.G.,Chang, Y.S. 대한생식의학회 1989 Clinical and Experimental Reproductive Medicine Vol.16 No.2

The purpose of this study was to examine the qualitative variation of human fetal-cord sera (HCS) and to accept the sera in human lVF-ET program. One hundred and sixteenth RCS were tested with 1772 2-cell embryos of F1 (C57BL x CBA) virgin mice, Ten to sixteenth embryos were cultured in m-KRB medium with a aliquot of each serum (10%, v/v) or with bovine serum albumin(O.4%, w/v) as a control medium. Embryonic development were recorded at every 24hr for 4 days by such events as cellular compaction, cavitation, and hatching. In the control groups of eight assays, 98.1%(106/ 108) of 2-ce1l embryos developed above expanded blastocyst and the embryonic development was unified through the tests. But the developmental pattern in medium with each serum was various. Namely, the sera that supported development of 100% 2-cell embryos to above morula, early blastocyst, expanded blastocyst and hatching blastocyst was 45,7%(53/116) , 35.3%(41/116), 15.5%08/116.) and 6.9-%(8/116), respectively. And the sera that supported development of above 80% 2-cell embryos to the each embryonic stage was 92.2% (107/116), 83.6%(97/116), 63.8%(74/116) and 36.2%(42/116), respectively. Meanwhile two kinds of toxic pattern to the embryonic development were observed in some sera. The first pattern is that some sera arrested development of most embryos in pre- or post-stage of morula or blastocyst. The second pattern is that some sera promoted or arrested a part of embryos in the same dish. The ability of serum was depended on the batch of serum. Finally we could accept 69%(80/116) of the tested sera for human IVF-ET program. The base line for acceptance was the ability that supported above 80% 2-ce1l embryos to blastocyst. But some deterious sera were contained in this range. We cut off about 10% of the sera (83.6% , 97/116) that passed the baseline. This final percent of sera was similar to that of grade N of this study.

원발성 무월경환자의 세포유전학적 연구

문신용,차상헌,이진용,장윤석,Moon, S.Y.,Cha, S.H.,Lee, J.Y.,Chang, Y.S. 대한생식의학회 1985 Clinical and Experimental Reproductive Medicine Vol.12 No.2

This investigation represents a cytogenetic study of 382 women with primary amenorrhea. 235 cases (61.5%) showed normal karyotype, 46, XX and l47 cases (38.5%) showed chromosomal abnormalities. Of 144 females with cytogenetic abnormalities, 44 cases (30.0%) showed a 45, X constitution: 37 cases (25.2%) with 45, X/46, XX: 3 (2.0%) 45, X/47, XXX: 11 (7.5%) 45, X/46, XX/47, XXX: 1 (0.7%) 45, X/46, XY: 3 (2.0%) 46, $XX_{p-}$: 2 (1.4%) 46, $XX_{q-}$: 6 (4.1%) 46, X, i ($X_q$): 1 (0.7%) 45, X/46, $XX_{q+}$: 1 (0.7%) 45, X/46, XX, inv(X): 1 (0.7%) 45, X, inv (9)/47, XXX, inv(9). 36 women (24.5%) had a 46, XY constitution and 1 case (0.7%) had a 47, XXX constitution.

마우스 난자(卵子)의 체외수정(體外受精)에 관(關)한 연구(硏究)

임용택,최승헌,김정구,문신용,이진용,장윤석,Lim, Y.T.,Choi, S.H.,Kim, J.G.,Moon, S.Y.,Lee, J.Y.,Chang, Y.S. 대한생식의학회 1984 Clinical and Experimental Reproductive Medicine Vol.11 No.2

The success of human in vitro fertilization (IVF) & embryo transfer (ET) has focused attention on the culture conditions that can provide optimal development of the preimplantation embryo. Studies of in vitro fertilization using mouse have direct implications to human IVF, since similar conditions are used for both species. Mouse IVF as a quality control system for human IVF & ET was studied since Feb., 1984. The results were as follows: 1. Egg retrieval following superovulation in IeR mice was l5.1${\pm}$5.3 eggs ovulated/mouse (Mean${\pm}$ S.D.) 2. In vitro cleavage rate was 61.7% (1146 eggs cleaved/l858 eggs inseminated) and % blastocyst was 42.6%. 3. In comparison with two media of Ham's F-10 and m-KRB, in vitro cleavage rate were 40.9%/63.l% and %blastocyst were 44.3%/61.2% (P<0.05). 4. It was concluded that mouse IVF system has a valuable place in human IVF & ET as a quality-control system and in human reproductive physiology as a research model.

Clomiphene Citrate를 이용한 배란유도에 관한 연구

문신용,윤병구,윤보현,김정구,이진용,장윤석,Moon, S.Y.,Yoon, B.K.,Yoon, B.H.,Kim, J.G.,Lee, J.Y.,Chang, Y.S. 대한생식의학회 1985 Clinical and Experimental Reproductive Medicine Vol.12 No.2

Therapy for anovulation represents one of the more gratifying and successful type of infertility management. Despite the introduction of bromocriptine, human menopausal gonadotropin (HMG), and gonadotropin releasing hormone (GnRH), clomiphene citrate remains the mainstay of ovulation induction therapy. There is wide variability in reported rates of ovulation induction (57-91%) and conception (25-43%) following clomiphene therapy. Factors contributing to this variability among different reports are the differences in dosage and duration of therapy, different criteria utilized in selecting patients for clomiphene therapy and different luteal phase parameters of presumptive ovulation. A review of recent experience with clomiphene citrate in the Department of Obstetrics and Gynecology in Seoul National University from January, 1983 to May, 1985 yields the following conclusions: 1. Ovulation rate per total patients treated was, 69.0% 2. Pregnancy rate per total patients treated was 31.7%, and that per total patients ovulated was 45.9%. 3. Ovulation rate at the dosage level up to 150 mg/day (50.3-53.8%) was somewhat higher than that at 200 mg/day or more (33.3-34.6%), and pregnancy rate per total patients treated was comparable at each dosage level. 4. Ovulation rate per total patients ovulated at each dosage level, where ovulation and conception occurred, showed a decreasing tendency as the dosage increased, but pregnancy rate per total patients conceived was comparable except at 200 mg/day. 5. Cumulative pregnancy rate per total patients conceived in each ovulatory cycle was 68.9% in 3 cycles, 88.9% in 4 cycles, and 100% in 6 cycles.

생쥐 모델을 이용한 배아의 할구 생검법과 할구가 생검된 배아의 배양시 공배양 효과에 관한 연구: 인간에서의 착상 전 유전진단 기술 개발을 위한 동물실험 모델의 개발

김석현,류범용,지병철,최성미,김희선,방명걸,오선경,서창석,최영민,김정구,문신용,이진용,채희동,김정훈,Kim, S.H.,Ryu, B.Y.,Jee, B.C.,Choi, S.M.,Kim, H.S.,Pang, M.G.,Oh, S.K.,Suh, C.S.,Choi, Y.M.,Kim, J.G.,Moon, S.Y.,Lee, J.Y.,Chae, H.D.,Kim, C.H. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.

난자공여 프로그램에서 난자수혜자의 연령이 임신율에 미치는 영향에 관한 연구

서창석,오선경,김석현,최영민,김정구,문신용,이진용,Suh, C.S.,Oh, S.K.,Kim, S.H.,Choi, Y.M.,Kim, J.G.,Moon, S.Y.,Lee, J.Y. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.2

Oocyte donation program developed to reach the pregnancy in those patients suffering from premature ovarian failure or surgery induced menopause, particularly in their reproductive age. With technical advances and popularity of ART (assisted reproductive technology), the indication of oocyte donation program extended to low responders, and even to naturally menopaused patients that has led them quite successfully to getting in pregnancy. The purpose of this study was to evaluate which one is involved in the decline of fertility between the oocyte and uterine factor. One hundred five cycles of oocyte donation program were performed in 84 patients from Jan., 1993 to Dec., 1996. Oocytes were donated from healthy, young, fertile anonymous donors or relatives or infertile patients with supernumerary oocytes. The study population was divided into 3 groups according to the age of recipients. Group 1 was less than 35 years old, Group 2 was between 35 to 39 years old, and Group 3 was more than 39 years old. The results were as follows: The mean age of oocyte donor was $31.5{\pm}3.3$ (range; 25-36). The mean concentration of basal serum FSH and peak serum estradiol were not different among groups. The mean number of oocytes retrieved from donors, embryos transferred to recipients, and fertilization rate were not different among groups. The clinical pregnancy rate was 37.3% in Group 1, 31.6% in Group 2, and 31.6% in Group 3, respectively. The spontaneous abortion rate was 16.0% in Group 1, 16.7% in Group 2, and 16.7 in Group 3, respectively. The multiple pregnancy rate was 20.0% in Group 1, 16.7% in Group 2, 16,7% in Group 3, respectively, The implantation rate was 11.3% in Group 1, 10.3% in Group 2 and 10.0% in Group 3, respectively. All of the pregnancy outcomes were not different statistically among groups. In conclusion, endometrial receptivity does not seem to be impaired as age increases with transfer of good quality embryos and adequate endometrial preparation.

동결보존시 생쥐 전핵배아의 시기에 따른 생존율과 발생율의 비교

김희선,류범용,오선경,서창석,김석현,최영민,김정구,문신용,이진용,Kim, H.S.,Ryu, B.Y.,Oh, S.K.,Suh, C.S.,Kim, S.H.,Choi, Y.M.,Kim, J.G.,Moon, S.Y.,Lee, J.Y. 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1

This study was designed to evaluate the influence of pronuclear age on the survival and post-thawing development after cryopreservation of mouse embryos. Freezing and thawing were performed in the different pronuclear stages of mouse embryos after IVF. Embryos were obtained from $F_1$ hybrid mice and classified into 4 groups according to the pronuclear stage (6hr, 9hr, 12hr and 15hr after insemination). Pronuclear ova were slowly cooled in a biological freezer using 1.5M 1,2-propanediol and 0.1M sucrose as cryoprotectant. Thawing was done at room temperature and 1,2-propanediol was removed by multi-step dilutions. Both frozen-thawed embryos and control fresh embryos were cultured in vitro in Ham's F-10 medium supplemented with 4mg/ml BSA. In control group, the development rate after 48hr was 99.3%, and the complete hatching rate after 144hr was 61.3%. In experimental groups, the survival rate after thawing was 95.4% in 6hr, 88.7% in 9hr, 75.2% in 12hr and 62.4% in 15hr after insemination, the development rate after 48hr was 61.1, 77.0, 67.0 and 79.6%, respectively, and the complete hatching rate after 144hr was 25.7, 43.7, 42.2 and 60.0%, respectively. The survival rate in 15hr was significantly lower (p<0.05) compared with other groups. In vitro development rates after 48hr were similar in all groups, but complement hatching rate was significantly lower (p<0.05) in 6hr group. In conclusion, cryopreservation of mouse pronuclear ova with 2 distinct pronuclei (9hr and 12hr groups) showed better results after thawing compared with early (6hr group) or late pronuclear ova just prior to cleavage (15hr group).