착상 전 유전진단 기술 개발의 동물실험 모델로서 할구 생검된 생쥐 배아에서 동결보존 융해 후 배아 발생 양상과 공배양 효과에 관한 연구

김석현,김희선,류범용,최성미,방명걸,오선경,지병철,서창석,최영민,김정구,문신용,이진용,채희동,김정훈,Kim, Seok-Hyun,Kim, Hee-Sun,Ryu, Buom-Yong,Choi, Sung-Mi,Pang, Myung-Geol,Oh, Sun-Kyung,Jee, Byung-Chul,Suh, Chang-Suk,Choi, Young-Min,Kim, Jung 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.1

Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

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.

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.

난자 세포질내 정자 주입술 후 동결보존 배아이식: 고식적 체외수정시술과의 비교 연구

김석현,지병철,정병준,김희선,류범용,방명걸,오선경,손철,서창석,최영민,김정구,문신용,이진용,Kim, S.H.,Jee, B.C.,Jung, B.J.,Kim, H.S.,Ryu, B.Y.,Pang, M.G.,Oh, S.K.,Shon, C.,Suh, C.S.,Choi, Y.M.,Kim, J.G.,Moon, S.Y.,Lee, J.Y. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

The objective of this study was to compare retrospectively the survival and pregnancy rates(PR) of cryopresered-thawed embryos obtained from intracytoplasmic sperm injection (ICSI) or conventional in vitro fertilization (IVF). Ninety-six cycles of cryopresered-thawed embryo transfer (ET) were performed in 79 patients from June, 1996 to September, 1997 and grouped as followings: 20 cycles (16 patients) inseminated by ICSI (ICSI Group) and 76 cycles (63 patients) by conventional IVF (IVF Group). Slow-freezing and rapid-thawing protocol was used with 1.5M propanediol (PROH) and 0.1M sucrose as cryoprotectant. All embryos were frozen-thawed at the two pronuclear (2 PN) stage excluding four cycles in which the early cleavage stage embryos were frozen, and allowed to cleave in vitro for one day before ET. The duration from freezing to thawing was comparable in both groups ($mean{\pm}SD$, $112.1{\pm}80.0$ vs. $124.8{\pm}140.1$ days). The age of female ($31.2{\pm}3.4$ vs. $32.6{\pm}3.3$ years) and the endometrial thickness prior to progesterone injection ($9.4{\pm}2.0$ vs. $9.3{\pm}1.8$ mm) were also comparable in both groups. There was no significant difference in the outcomes of cryopreserved-thawed ET between two groups: survival rate ($85.2{\pm}16.1%$ vs. $82.2{\pm}19.7%$), cleavage rate ($96.9{\pm}6.7%$ vs. $94.7{\pm}13.0%$), cumulative embryo score (CES, $54.5{\pm}31.1$ vs. $49.0{\pm}20.0$), preclinical loss rate (5.0% vs. 5.3%), clinical miscarriage rate (0% vs 29.4%), clinical PR per transfer (35.0% vs. 22.4%), implantation rate (9.9% vs. 5.6%), and multifetal PR (42.9% vs. 17.6%). In conclusion, human embryos resulting from ICSI can be cryopreserved-thawed and transferred successfully, and the survival rate and PR are comparable to conventional IVF.

특허 침해권(I)

김석현,Kim, Seok-Hyeon 한국광학기기산업협회 2006 光學世界 Vol.103 No.-

신라대종의 맥놀이 조절

김석현,이중혁,Kim, Seockhyun,Lee, Joong Hyeok 한국음향학회 2017 韓國音響學會誌 Vol.36 No.3

신라대종은 성덕대왕신종을 재현하려는 목적으로 제작 되었으며, 동일한 구조와 문양을 갖도록 복원되었다. 가장 어려운 문제는 성덕대왕신종의 소리에 걸 맞는 웅장하고 조화로운 타격음과 끊어질 듯 이어지는 여음의 맥놀이를 재현하는 것이었다. 특히 맥놀이는 주조과정에서 발생하는 제어 불가능한 미세 비대칭성에 기인하므로, 주조 전에는 그 특성을 예측하거나 제어할 수 없다. 본 연구에서는 주조 후 맥놀이의 조절 작업을 통하여 신라대종의 맥놀이가 강하고 적절한 주기를 갖도록 만드는 방법과 과정을 소개한다. 강한 맥놀이를 만드는 모드 쌍과 당좌의 배치 조건을 구하였고, 적절한 주기를 갖도록 종 내부의 두께를 국부적으로 감소시켰다. 이 작업은 등가 종을 이용한 시뮬레이션 결과에 따라 수행되었다. 그 결과 약하고 매우 길었던 초기의 맥놀이는 강하고 적절한 주기의 맥놀이로 만들어졌다. Silla Great Bell was made to reproduce King Seongdeok Divine Bell and it was restored to have the same structure and patterns. The most difficult problem was to reproduce the magnificent striking sound and dynamic hum tone with strong beat like in King Seongdeok Divine Bell. Especially, beating sound is attributed to the uncontrollable asymmetry occurring in the casting process, so it can not be predicted or controlled before casting. In this study, we introduce the method and process to make Silla Great Bell have a strong beat with a proper period. Position conditions of mode pairs and striking point for a strong beat were identified. Bell thickness was locally decreased to make proper period of beat. The process was performed according to the simulation result of an equivalent bell model. As a result, the original weak and long beat was made to a strong beat with a proper period.

축열 시스템 용어

김석현,강기호,김영인,최규명,Kim, Seok-Hyeon,Gang, Gi-Ho,Kim, Yeong-In,Choe, Gyu-Myeong 한국설비기술협회 1994 설비 : 공조ㆍ냉동ㆍ위생 Vol.11 No.3

2006년 하천정책 추진방향

김석현,Kim, Seok-Hyeon 한국하천협회 2006 하천과 문화 Vol.2 No.2

다태임신에서의 선택적 유산술시 복식 천자와 질식 천자의 비교 연구

김석현,문신용,이진용,Kim, S.H.,Moon, S.Y.,Lee, J.Y. 대한생식의학회 1996 Clinical and Experimental Reproductive Medicine Vol.23 No.1

The number of multifetal pregnancies has increased dramatically as a result of the widespread clinical use of ovulation induction and assisted reproductive technology(ART) in infertile patients. In multifetal pregnancies, the adverse outcome is directly proportional to the number of fetuses within the uterus, primarily because of an increased predisposition to premature delivery. It is extremely difficult to counsel patients about the expected outcome of pregnancies involving three or more fetuses. To increase the chances of delivering infants mature enough to survive without being irreversibly damaged by the sequelae of marked prematurity, selective fetal reduction(SFR) to the smaller number of fetuses should be considered in multifetal pregnancies. From January, 1991 to December, 1992, transabdominal SFR in multifetal pregnancies was performed in 22 patients including 13 triplet, 7 quadruplet, 1 quintuplet and 1 heptuplet pregnancies. Transabdominal SFR using intracardiac KCI injection and aspiration of amniotic fluid was carried out in 8-13 weeks of gestation. After procedure, 20 patients were remained as twin pregnancies, and 2 patients as triplet pregnancies. There have been 11 sets of twin delivery including 2 stillbirths, 2 sets of triplet delivery including 1 stillbirth, and 1 singleton delivery. Six cases were delivered after 37 weeks of gestation, 4 cases in 33 - 37 weeks, and 1 case in 30 weeks. Unfortunately, 3 stillbirths occurred in 20-24 weeks of gestation, and 4 cases were aborted. As 7 losses of pregnanancy including 1 case of septic abortion occurred, the delayed fetal loss rate was 38.9%(7/18) in transabdominal SFR. All babies born after 30 weeks of gestation were healthy, and no fetal anomaly directly related to the procedure was encountered. From July, 1993 to February, 1995, transvaginal SFR was performed in 20 patients including 15 triplet, 4 quadruplet and 1 quintuplet pregnancies. Transvaginal SFR using the same method as transabdominal SFR was carried out in 8-11 weeks of gestation. After procedure, 19 patients were remained as twin pregnancies, and 1 patient as singleton pregnancy. There have been 13 sets of twin delivery including 2 stillbirths, and 1 singleton delivery. Six cases were delivered after 37 weeks of gestation, 5 cases in 36-37 weeks, and 1 case in 30 weeks. Unfortunately, 2 still-births occurred in 20 weeks and 21 weeks of gestation, respectively, and 2 cases were aborted. As 4 losses of pregnancy including 1 case of septic abortion occurred, the delayed fetal loss rate was 25.0%(4/16) in transvaginal SFR. No fetal anomaly directly related to the procedure was encountered. It is suggested that transvaginal SFR could be performed more easily and earlier with the lower fetal loss rate as compared with transabdominal SFR. In conclusion, SFR is a rather safe and ethically justified procedure that may improve the outcome of multifetal pregnancies.

가스이용 냉방시스템의 원리(2)

김석현,Kim, Seok-Hyeon 한국설비기술협회 1992 설비 : 공조ㆍ냉동ㆍ위생 Vol.9 No.5