인간의 체외수정 및 배아이식술 관련 용어와 기록 표준화

전진현,이동률,박동욱,지희준,윤산현,강인수,이규섭,최영민,Jun, Jin-Hyun,Lee, Dong-Ryul,Park, Dong-Wook,Chi, Hee-Jun,Yoon, San-Hyun,Kang, Inn-Soo,Lee, Kyu-Sup,Choi, Young-Min 대한생식의학회 2008 Clinical and Experimental Reproductive Medicine Vol.35 No.2

보조생식술 센터에서 ISO 9001 : 2000 품질경영시스템의 도입 및 시행의 효용성

전진현,박용석,이형송,김순덕,황선희,한수경,김재호,송인옥,강인수,궁미경,Jun, Jin-Hyun,Park, Yong-Seog,Lee, Hyoung-Song,Kim, Soon-Duk,Hwang, Son-Hee,Han, Su-Kyoung,Kim, Jae-Ho,Song, In-Ok,Kang, Inn-Soo,Koong, Mi-Kyoung 대한생식의학회 2007 Clinical and Experimental Reproductive Medicine Vol.34 No.2

목 적: ISO 9001:2000 품질경영시스템은 국제적인 인중 기준에 의해 제품 및 서비스의 품질을 적절하게 판정하고 이를 향상시키기 위해 시행되고 있다. 본 연구에서는 이러한 품질경영시스템을 보조생식술 센터에 성공적으로 도입하고 적용하는 과정에서 확인된 효용성에 대해 기술하고자 한다. 연구방법: 제일병원 아이소망센터는 2004년 1월부터 ISO 9001:2000 인증을 위한 활동을 시작하였으며, 고객만족도조사와 주기적인 내부심사와 사후심사를 실시하였다. 심사과정에서 확인된 부적합 사항에 대한 시정 및 예방 조치와 이에 따른 효과성을 검증하였으며, 고객불만사항에 대한 지속적인 관리와 개선을 위한 프로젝트를 진행하였다. 결 과: 본 센터는 초일류 불임센터로 성장하고 최적의 품질경영시스템을 확립하기 위한 품질방침을 설정하여, 2004년 6월에 한국품질재단 (Korean Foundation for Quality)으로부터 "체외수정 및 배아이식 시술에 관련된 연구"에 대하여 ISO 9001:2000 인증을 받았다. 이러한 품질방침을 실현하기 위해 품질경영시스템에 적합한 품질매뉴얼, 프로세스, 절차서, 지침서 등에 대한 문서화를 완료하였다. 삼 년 동안의 내부심사와 사후심사에서 각각 140건과 7건의 부적합이 확인되었으며, 이에 대한 시정조치를 실시하였다. 결 론: 본 센터에서는 ISO 품질경영시스템의 도입과 운영을 통해 고객만족도의 향상, 체계적인 문서화를 통해 업무의 투명화와 효율화가 가능하였다. 보조생식술 센터에서의 ISO 품질경영시스템은 보조생식술에 대한 국가적인 관리시스템을 효율적으로 운영하기 위한 기본적인 기관별 경영시스템으로서 활용이 가능할 것으로 생각된다. Objective: A quality management system of international standardization organization (ISO) 9001:2000 has been proposed to properly evaluate and improve the quality of productions and services. The purpose of this article was to describe on successful introduction and application of the ISO system to human assisted reproductive technology (ART) center. Methods: Center for reproductive medicine and infertility of Cheil General Hospital started the action for ISO 9001:2000 certification at January-2004. The ISO system of our center was monitored by measuring the customer satisfaction index and periodical internal and external audit. Reports of non-conformity described corrective and preventive actions for problems and occurrences that were not consistent with the standard process and operation of our center. Continuous management and improvement were performed on the project of customer dissatisfaction. Results: Our center has got the certification of ISO 9001:2000 for "Research and development of IVF-ET program for infertility treatment" from Korean Foundation for Quality at June-2004. The policy of ISO was "To establish the best center of reproductive medicine and infertility" and "To establish the best quality management system". We found 140 cases and 7 cases of non-conformity in internal and external audit for three years, respectively, and performed corrective action. Conclusion: Improvement of customer satisfaction, systemic documentation and transparency and efficacy of working procedure were achieved by application and operation of the ISO system to our center. This ISO system could be used as a basic institutional management system for the national guideline to the human ART center.

Effect of Hydrosalpingeal Fluid on the Implantation in-vitro in a Murine Model

전진현,궁미경,임천규,김수경,강인수,Jun, Jin-Hyun,Koong, Mi-Kyoung,Lim, Chun-Kyu,Kim, Soo-Kyung,Kang, Inn-Soo The Korean Society for Reproductive Medicine 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.2

연구목적:인간의 체외수정 및 배아이식술에서 난관수종을 갖는 환자에서 임신율과 착상률이 감소된다는 보고들이 있지만 이에 대한 명확한 기작은 밝혀지지 않았다. 본 연구에서는 생쥐 배아를 이용한 체외 착상모델에서 인간의 난관수종액(HSF)이 착상과정에 미치는 영향을 알아보고자 하였다. 연구재료 및 방법 난관수종액은 난관수종으로 수술을 받은 8명의 환자로부터 채취하였으며, 실험에 사용하기 전까지 냉동고에 보관하였다. 생쥐의 포배기 배아는 2-세포기배아를 3일 동안 배양하여 그 중 상태가 양호한 포배기 배아만을 선별하여 투명대를 제거한 후 사용하였다. 기본 배양액으로는 Ham's F-10을 사용하였으며, 배양 시 기본 배양액만을 사용한 경우를 group Ⅰ으로 하였고, 기본 배양액에 0.5% FBS를 첨가한 경우를 group Ⅱ, 0.5% FBS와 50% HSF를 첨가한 경우를 group Ⅲ , 100% HSF에 0.5% FBS를 첨가한 경우를 group Ⅳ,100% HSF만을 사용한 경우를 group Ⅴ로 하였다. 투명대를 제거한 포배기 배아를 각각의 HSF에 대한 5종류의 배양액에서 48시간 동안 배양하였다. 체외 착상 유무는 부착 부위에서 크기가 커진 영양세포들을 관찰하여 판정하였으며, 착상 부위의 표면적은 화상분석기를 이용하여 산출하였다. 결 과: 생쥐 배아의 체외 착상률은 group Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 0%, 98.9%, 77.5%, 40.4%, 10.0%로 나타났으며, 착상 부위의 평균 표면적은 group Ⅱ, Ⅲ, Ⅳ, Ⅴ에서 각각 $74,675{\pm}25,201{\mu}m^2$, $59,024{\pm}25,877{\mu}m^2$, $45,156{\pm}22,654{\mu}m^2$, $38,254{\pm}17,115{\mu}m^2$이었다. 체외 착상률과 부위의 표면적은 HSF의 농도가 증가함에 따라 통계적으로 유의하게 감소하였다(p<0.001). 결론:인간의 난관수종액(HSF)은 생쥐 배아의 체외 착상과 영양배엽세포의 증식을 억제하는 것으로 확인되었으며, 이러한 원인이 난관수종을 갖는 환자에서 임신율이 낮은 것과 밀접한 관련이 있을 것으로 생각된다.

고환조직 정자채취술 (TESE)의 정자 상태에 따른 세포질내 정자주입술의 (ICSI)수정률과 임신율

전진현,김정욱,박용석,이호준,서주태,이유식,손일표,전종영,Jun, Jin-Hyun,Kim, Jeong-Wook,Park, Yong-Seog,Lee, Ho-Joon,Seo, Ju-Tae,Lee, Yu-Sik,Son, Il-Pyo,Jun, Jong-Young 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

In IVF-ET program, intracytoplasmic sperm injection(ICSI) has been performed with testicular sperm extraction(TESE) in case of no normal spermatozoon could be retrieved from the epididymis. We wished to see whether the quality of testicular sperm affect the fertilization and pregnancy rate in TESE-ICSI cycles(n=40). These cycles were classified into three groups by the total number of normal motile spermatozoa(TNMS) in the TESE sample: i) good sperm(GS) group(n=12), TNMS > 10,000; ii) moderate sperm(MS) group(n=19), 1,000 < TNMS < 10,000; iii) poor sperm(PS) group(n=9), TNMS < 1,000. Among 423 injected oocytes, 307(72.6%) oocytes were normally fertilized and 43 zygotes were cryopreserved. The fertilization rates of GS group(79.3%) and MS group(75.9%) were significantly(p<0.005) higher than PS group(60.2%). After the embryo transfer(n=40), clinical pregnancy was obtained in 14 cycles(35.0%) and on-going pregnacy in 13 cycles(32.5%). The clinical and on-going pregnancy rates were similar in each group. From these results it can be concluded that testicular spermatozoa are successfully used with ICSI in IVF-ET program in spite of very poor quality of TESE sample.

세포질내 정자주입을 시행한 인간 난자의 전핵 형성 및 배아 발생에 관한 연구

전진현,임천규,박용석,이호준,김종흡,김문규,Jun, Jin-Hyun,Lim, Chun-Kyu,Park, Yong-Seog,Lee, Ho-Joon,Kim, Jong-Heup,Kim, Moon-Kyoo 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.3

Mammalian, including human, spermatozoa undergo morphological and physiological changes during sperm maturation. There were, these changes may affect the fertilization and embryonic development. In this study, we examined the pronucleus formation, pronucleus disappearance and embryonic development in the human oocytes fertilized by intracytoplasmic sperm injection (ICSI). The injected spermatozoa were grouped into ejaculated, epididymal and testicular by the collecting region. Among 363 metaphase II injected oocytes, 287(79.1%) oocytes were normally fertilized and displayed two pronuclei. There were no difference in the fertilization rates and in the pronucleus formation and pronucleus disappearance at 16, 20 and 24 hr after ICSI, among the each spermatozoa group. Also, at 64 hr, the appearance of embryonic development was similar. From these results it can be concluded that there was no difference of maturity among the sperm collected from ejaculated, epididymis and testis in the pronucleus formation and embryonic development. Therefore, testicular spermatozoa are successfully used with ICSI in IVF-ET program.

체외수정 및 배아이식술에서 세포질내 정자주입술(ICSI)의 수정률과 임신율

전진현,이호준,김정욱,박용석,이유식,홍재엽,손일표,전종영,Jun, Jin-Hyun,Lee, Ho-Joon,Kim, Jeong-Wook,Park, Yong-Seog,Lee, Yu-Sik,Hong, Jae-Yup,Son, Il-Pyo,Jun, Jong-Young 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.3

Intracytoplasmic sperm injection(ICSI) was known as effective method in treatments of couples who unable to be helped by conventional in vitro fertilization. In 78 treatment cycles of 78 infertile couples using ICSI performed at our infertility clinic between May and August 1994 were analyzed. These patients were classified two groups, andrological factor(AF) and non-andrological factor(non-AF) group. The AF group, which had abnormal sperm physiology, included oligozoospermia, asthenozoospermia, oligoasthenoteratozoospermia(OATS) and microsurgical epididymal sperm aspiration(MESA) patients. The non-AF group, which had abnormal oocyte physiology, included abnormal zona pellucida, poor quality of oocyte and immune factor infertile patients. A single spermatozoon was injected into the ooplasm of 776 metaphase II oocytes. The fertilization rate was 44.6%(346/776) and 319 embryos were transferred. After 73 embryo transfers(93.6% of treatment cycles) 23 pregnancies were estabilshed, i. e. pregnancy rate of 29.4% per started cycle and 31.5% per embryo transfer. Fertilization rate of AF and non-AF group was 46.2% and 35.8%, pregnancy rate was 34.5%(20/58) and 20.0%(3/15), respectively. In order to increase the pregnancy rate, assisted hatching(AHA) has done after lCSl in 47 treatment cycles. Pregnancy rate of ICSI with AHA and without AHA group was 34. 0% (16/47) and 26.9%(7/26), respectively. ICSI was more effective in andrological factor infertility and the pregnancy rate was increased by ICSI with AHA procedure.

생리혈에 존재하는 탈락된 자궁내막조직의 체외배양 시 나타나는 변화와 특성에 관한 연구

전진현(Jin Hyun Jun),궁미경(Mi Kyoung Koong),강인수(Inn Soo Kang),양광문(Kwang Moon Yang),홍수정(Soo Jeong Hong),김문규(Moon Kyoo Kim) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.1

목적: 자궁내막증의 체외 모형에 적합한 생리혈 내의 탈락된 자궁내막조직을 체외배양하면서 나타나는 변화와 특성에 관하여 살펴보고자 하였다. 연구 방법: 정상적인 생리주기를 갖는 10명의 여성으로부터 Wallace catheter를 이용하여 생리혈을 채취하였다. 원심분리 방법으로 생리혈을 세척한 후, 수획한 탈락된 자궁내막조직은 10% fetal bovine serum이 첨가된 Ham's F-10배양액에서 7일 동안 배양하였다. 배양한 자궁내막조직 세포의 특성을 확인하기 위해 cytokeratin과 vimentin에 대한 항체를 이용하여 면역조직학적 염색을 실시하였다. 결과: 생리혈의 평균 양은 0.7ml이였으며, 세척 후 혈액세포와 탈락된 자궁내막조직이 포함된 침전부위의 평균 양은 0.3ml 이였다. 체외배양 한 자궁내막조직의 15% 정도가 배양접시 표면에 부착된 후 증식하였다. 일차적으로 상피세포와 섬유아세포가 배양접시의 표면에 부착하여 증식하였으며, 기질세포들은 상피세포 위에서 증식하였다. 면역조직학적 염색에서 상피세포, 기질세포, 섬유아세포 등이 확인되었으며, 공촛점주사 현미경을 이용한 체외배양된 탈락된 자궁내막조직의 3차원적인 구조 관찰에서 외부에는 상피세포가, 내부에는 기질세포가 존재함을 확인하였다. 결론: 생리혈 내의 탈락된 자궁내막조직의 체외배양 시 상피세포와 기질세포가 밀접한 연관성을 유지하면서 증식하는 것을 관찰하였으며, 이러한 체외배양 체계는 자궁내막증의 병인에 대한 연구에 유용하게 적용될 수 있는 체외 모형으로 사료된다. Objective: To evaluate the viability and the characteristics of shed endometrial tissues obtained from menstrual fluid during in-vitro culture. Methods: The menstrual fluids were collected using Wallace catheter from uterine cavity in 10 women with regular menstruation. The menstrual fluids were washed twice, and the pellets, containing blood cells and shed endometrium, were collected and diluted fivefold with Ham's F-10 medium containing 10% fetal bovine serum. The cell suspension was placed on culture dishes, and cultured for 7 days in an incubator. To evaluate the characteristics of the cultured endometrial cells, immunohistochemical (IHC) staining was performed using anti-cytokeratin and anti-vimentin antibody. Results: The mean volume of menstrual fluids and pellets were 0.7ml and 0.3ml, respectively. Only 15% of the shed endometrial tissues were attached and proliferated in culture dishes, which was considered to have viability. Initially, endometrial epithelial cells and fibroblasts were attached and proliferated, and the area of these cells was increased according to prolong the culture time. Stromal cell colonys were located and proliferated on the epithelial cells. IHC staining showed strongly positive for cytokeratin in epithelial cells and for vimentin in stromal cells. In the confocal microscopic observation of 3-dimensional structure of cultured endometrium, cytokeratin-positive cells (epithelial cells) were located in the pheriphery and cytokeratin-negative cells (stromal cells) inside of the structure. Conclusion: From our study, shed endometrial tissues in menstrual fluid showed meaningful viability and closed relationship between epithelial cells and stromal cells during in-vitro culture. Thus, we suggest that the in-vitro culture system of shed endometrium is a suitable model for researches of endometriosis.

균형 전좌 또는 Robertsonian 전좌 보인자의 체외수정 및 배아이식술에서 형광직접보합법을 이용한 착상전 유전자진단의 임상적 적용

전진현(Jin Hyun Jun),송견지(Gyun Jee Song),김정욱(Jeong Wook Kim),박소연(So Yeon Park),김계현(Kye Hyun Kim),최범채(Bum Chae Choi),궁미경(Mi Kyoung Koong),강인수(Inn Soo Kang),임천규(Chun Kyu Lim),한미현(Mi Hyun Han) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.7

목적 : 본 연구는 균형 전좌 또는 Robertsonian 전좌 보인자의 체외수정 및 배아이식술에서 형광직접보합법을 이용한 착상전 유전자진단을 시행하여 그 결과와 효율을 알아보고자 시행하였다. 연구방법 : 본원에서 착상전 유전자진단을 시행한 15쌍, 25주기의 체외수정 및 배아이식술을 연구대상으로 하였으며, 3주기에서 54개의 극체를 이용하여, 22주기에서 234개의 할구를 이용하여 형광직접보합법으로 염색체의 숫적 이상을 분석하였다. 형광직접보합법을 이용하여 분석한 후 정상의 형광직접보합법 signal을 나타내는 배아만을 모체에 이식하였다. 임신이 확인된 경우 양수천자의 방법으로 태아의 핵형을 확인하였다. 결과 : 극체를 이용한 형광직접보합법 분석에서, 18개의 극체가 정상이었고, 3주기 모두에서 배아를 이식하였으며, 형광직접보합법의 효율은 95.0%였다. 할구를 이용한 형광직접보합법 분석에서는 49개의 배아가 정상으로 확인되었다. 정상의 배아를 확인할 수 없었던 1주기를 제외한 21주기에서 배아이식을 시행하였으며, 형광직접보합법의 효율은 92.7%였다. 3주기에서 임신이 되었고, 2주기에서 건강한 균형 전좌 보인자인 남아와 Robertsonian 전좌 보인자인 여아가 태어났다. 1주기는 임신이 진행중이며, 양수천자에서 정상의 핵형으로 확인되었다. 결론 : 형광직접보합법을 이용한 착상전 유전자진단은 균형 전좌 또는 Robertsonian 전좌 보인자의 체외수정 및 배아이식술에 성공적으로 적용되어 태아의 염색체 이상으로 인하여 발생되는 여러가지 문제를 해결할 수 있는 효과적인 방법으로 사료된다. Objective : This study was performed to evaluate the efficiency of preimplantation genetic diagnosis (PGD) using fluorescence in-situ hybridization (FISH) in Robertsonian or balanced reciprocal translocation carriers in human IVF-ET programm.Method : FISH was carried out in 25 cycles of 15 couples. Two-color FISH analysis was performed on 54 polar bodies in 3 cycles and 234 blastomeres in 22 cycles. After FISH analysis, the embryos with normal FISH signals were transferred into mother's uterus.Results : In FISH analysis of polar bodies, 18 nuclei of polar bodies were normal and 12 embryos were transferred in 3 cycles. FISH efficiency per oocyte was 95.0% in cases using polar bodies. In FISH analysis of blastomeres, 49 embryos were normal and transferred in 21 cycles. FISH efficiency per embryo was 92.7% using blastomeres. At present, three pregnancies were achieved. A girl and a boy were delivered. Both of them were translocation carriers. The other conceptus showed normal karyotype.Conclusion : According to this study, PGD using FISH can be successfully applied for the patients with translocations of chromosomes.

배양액 내의 마그네슘 이온이 생쥐 초기 배아 발생에 미치는 영향

최수진,전진현,박용석,배인하,Choi, Soo-Jin,Jun, Jin-Hyun,Park, Yong-Seog,Bae, In-Ha 대한생식의학회 2001 Clinical and Experimental Reproductive Medicine Vol.28 No.3

Objective: The present study was undertaken to examine the effects of magnesium ion in the culture medium on the development of mouse fertilized oocytes either before or after pronuclear formation, and to investigate whether the effect of magnesium ion is related with the redistributional change of mitochondria. Methods : Fertilized oocytes obtained from the oviducts of mice at 15 hr after hCG injection before pronuclear formation (pre-PN) or 21 hr after hCG injection after pronuclear formation (post-PN) were used. The embryos were cultured for 3 days with basic T6 medium-magnesium free and various concentrations of magnesium ion, 0.0, 0.5, 1.0, 2.0, 4.0 or 8.0 mM, respectively. After culture, the developmental stages of embryos and the number of nuclei were evaluated. To observe the effects of magnesium ion on the mitochondrial distribution, fertilized oocytes were collected at 21 hr after hCG injection and cultured for 6 hr with various concentration of magnesium ion. As a control, fertilized oocytes with pronuclei at 27 hr after hCG injection were used. Results: The concentration of magnesium ion to accelerate the in vitro development of mouse fertilized oocytes appeared to be at 2.0 mM for the pre-PN and the post-PN stage embryos. In the mitochondrial redistribution patterns, the embryos cultured in 2.0 mM concentration of magnesium ion showed the highest percentage (22.6%) of distinct perinuclear clustering pattern comparing to other experimental group. Conclusion: The effect of magnesium ion may be related to the cytoplasmic redistribution of mitochondria. This relationship seems to connect the developmental competence of preimplantation mouse embryos in vitro. These results can suggest that higher concentration of magnesium ion (2.0 mM) than those of conventional culture medium ($0.2{\sim}1.2\;mM$) is more suitable for in vitro culture of preimplantation mouse embryos.

불임여성에서 NAT2, GSTM1, CYP1A1 유전자 다형성과 자궁내막증의 상관관계에 관한 연구

송현정,전진현,최혜원,허걸,강인수,궁미경,이형송,Song, Hyun-Jeong,Jun, Jin-Hyun,Choi, Hye-Won,Hur, Girl,Kang, Inn-Soo,Koong, Mi-Kyoung,Lee, Hyoung-Song 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.2

Objective: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. Materials and Methods: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. Result: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. Conclusion: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.