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.

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.

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.

쉽게 배우는 특허 Q&A - 산업재산권

김석현,Kim, Seok-Hyeon 한국광학기기산업협회 2011 光學世界 Vol.134 No.-

만경강에서 하천정비에 의하여 격리된 구하도의 어류 군집 구조: 연결성 복원을 위한 제안

김석현,전형태,조강현,Kim, Seog Hyun,Cheon, Hyoung Tae,Cho, Kang-Hyun 응용생태공학회 2015 Ecology and resilient infrastructure Vol.2 No.1

본 연구는 하천에서 격리된 구하도와 본류에서 어류 군집 구조의 차이를 비교하기 위하여, 만경강에서 2곳의 본류와 단절된 구하도를 선정하여 환경 요인과 어류 군집 구조를 조사하였다. 측정된 환경 자료를 이용하여 주요인분석을 실시한 결과, 구하도는 하상이 주로 진흙과 모래로 이루어졌고 식생 피도가 높은 반면 본류는 수로 폭이 넓고 하상이 왕자갈, 잔돌, 호박돌 등 다양한 하상구조를 보였고 용존산소 및 전기전도도가 높게 나타났다. 어류 자료를 사용하여 군집분석 결과, 구하도 군집과 본류 군집으로 뚜렷이 구분되었다. 구하도는 호소 및 소하천에 서식하는 어류 군집 구조가 나타났고, 본류는 강과 같은 대형하천에 서식하는 어류군집 구조를 나타냈다. 비모수다차원척도법 결과에 의하면 구하도에 분포하는 어류는 식생 피도와 높은 양의 상관관계를 가진 반면 본류에 분포한 어류는 용존산소, 전기전도도와 높은 상관관계를 보였다. 본 연구 결과, 구하도는 본류와의 연결성이 단절되면서 본류와 비교할 때 어류 서식 환경이 완전히 달라짐으로써 어류 군집 구조가 큰 차이를 보이고 어류의 산란처 및 은신처 기능도 상실한 것으로 판단되었다. 따라서 하천의 어류 생물다양성 증진을 위하여 구하도의 횡적 연결성을 복원하기 위한 노력이 필요하다. This study investigated the difference in fish community structures in a main channel and an isolated former channel, considering the environmental factors in the Mangyeong River, Korea. Principal component analysis (PCA) with environmental factors showed that former channels were composed of a fine substrate covered by in-stream vegetation, whereas the main channel was covered by a wide range of substrates with a higher dissolved oxygen and conductivity. The result of the hierarchical cluster analysis with species abundance delineated to the four main groups; three abandoned channel groups and one main channel group. Non-metric multidimensional scaling (NMDS) showed that fish community structures of each study site differed from environmental factors: former channel fish communities were positively related to in-stream vegetation cover, whereas main channel fish communities were positively associated with dissolved oxygen and conductivity. The results indicated that channelization, where there was a separation between the former channel and the main channel, had detrimental effects on fish community structures of both the main channel and the abandoned channel in the Mangyeong River. In conclusion, this study suggested that the connectivity between the main channel and abandoned channel were required to enhance both habitat structural diversity and species diversity of the Mangyeong River.

한국의 조립가공 · 부품 · 소재 · 장비산업, downstream에서 upstream으로 도약 시동

김석현,Kim, Seok-Hyeon 한국광학기기산업협회 2006 光學世界 Vol.101 No.-

최근 무역흑자기조가 유지되고 있으며 2003년 이후 수출증가율이 수입증가율을 앞서나가고 있어 무역 흑자 규모가 2004년 약 200억불로 커지고 있다. 그러나 주요기간산업의 대일의존은 여전하다. 특히 광학산업을 포함한 기계산업은 역조규모가 가장 크며 대부분의 품목들이 대일수입에 의존하고 있고 그 뒤를 전자전기산업이 따르고 있다. 이처럼 한국의 자본재 산업은 해외에 의존하고 가공/조립을 통한 범용제품(후방산업) 양산에 치우쳐왔다. 따라서 기술개발능력이 있는 중소기업의 원천기술에의 도전을 적극적으로 지원하는 연구개발 사업이 필요하며, 기업이 보유한 특허와 같은 지적재산에 투자하는 지적재산캐피탈(IPC)을 도입하여 기술보유기업의 성장과 신기술의 시장진입을 도와야 할 것이다.

2차원 어류 서식처 모의를 이용한 격리된 구하도의 연결성 복원에 따른 서식지 개선 평가

김석현,김다나,조강현,Kim, Seog Hyun,Kim, Dana,Cho, Kang-Hyun 응용생태공학회 2015 Ecology and resilient infrastructure Vol.2 No.2

하천 본류와 구하도 사이의 횡적 연결성은 하천-홍수터 생태계에서 생태적 기능을 유지하는데 매우 중요하다. 본 연구는 2차원 평균수심 유한요소 모형인 River2D를 사용하여 만경강에서 직강화로 인해 격리된 구하도의 연결성을 가상으로 복원하고 평수기와 홍수기 조건에서 어류 서식처 변화를 평가하였다. 평가 대상 어종은 정수성 어종인 붕어 (Carassius auratus)와 유수성 어종인 피라미 (Zacco platypus)를 선정하였다. 물리서식처모의 결과에 의하면, 연결성 복원 전후의 붕어와 피라미의 가중가용면적 (weighted usable area, WUA)은 복원 후에 증가하였으며 양방향 복원이 일방향 복원보다 어류 서식처 복원에 효과적이었다. 또한 가중가용면적의 증가율은 평수기보다 홍수기에 더 높았다. 특히 피라미의 경우 홍수기에 구하도의 연결성이 양방향 복원되었을때 구하도 내의 가중가용면적이 복원 전 대비 약 4배로 크게 증가할 것으로 예측되었다. 본 연구 결과 만경강에서 본류와 구하도의 연결성이 복원되었을 때 정수성 어류와 유수성 어류의 서식처가 모두 증가할 것으로 예상되며, 일방향 복원보다 양방향 복원이 더 효과적인 것으로 나타났다. 또한 복원된 구하도는 홍수기 때 어류의 피난처 역할을 할 것으로 기대되었다. Lateral connectivity between a main channel and a former channel plays an integral role in maintaining ecological functions of stream-floodplain ecosystems. This study virtually restored the connectivity of the former channel, which is currently isolated by channelization, in the Mangyeong River, Korea. Fish habitat improvement after the connectivity restoration was evaluated using River2D, two-dimensional depth-averaged hydraulic modeling, depending on normal and flood flow conditions. Target fish species were crucian carp (Carassius auratus), which are known as lentic species, and pale chub (Zacco platypus), known as lotic species. The weighted usable area (WUA) of the two species was increased after the connectivity restoration: the two-way connection between the main and formal channels was more effective than the one-way connection. The result of the physical habitat simulation at a flood flow condition demonstrated an increased rate of the WUA than during a normal flow condition. In particular, the WUA of pale chub increased about four times on the two-way connectivity restoration. This result suggests that habitat availability of both lentic and lotic fish species will increase after a connectivity restoration, and a two-way connectivity restoration may be more effective. In addition, the restored formal channel would function as a shelter for fish during the flood season.

쉽게배우는 특허 Q&A - 우선심사

김석현,Kim, Seok-Hyeon 한국광학기기산업협회 2011 光學世界 Vol.133 No.-

친환경적 하천정비를 위한 법.제도적 방향

김석현,Kim, Seok-Hyeon 한국하천협회 2007 하천과 문화 Vol.3 No.2