Comparative Results of Embryo Development and Clinical Pregnancy using Sperm Retrieved from Fresh and Frozen-thawed Testicular Tissue from Patients with Obstructive and Non-obstructive Azoospermia

박용석,최수진,이선희,박동욱,임천규,전진현,궁미경,박찬우,송인옥,서주태,Park, Yong-Seog,Choi, Su-Jin,Lee, Sun-Hee,Park, Dong-Wook,Lim, Chun-Kyu,Jun, Jin-Hyun,Koong, Mi-Kyoung,Park, Chan-Woo,Song, In-Ok,Seo, Ju-Tae The Korean Society for Reproductive Medicine 2009 Clinical and Experimental Reproductive Medicine Vol.36 No.4

Objective: To compare the embryonic development and pregnancy results using sperms retrieved from fresh and frozen-thawed testicular tissue in patients with obstructive (OA) and non-obstructive azoospermia (NOA). Methods: A total two hundred twenty-two cycles of TESE-ICSI were performed in OA and NOA. Sperms were retrieved from fresh and frozen-thawed testicular tissue. ICSI was performed patient's own sperm. Fertilization was assessed 16~18 hrs after ICSI. Embryo development and pregnancy rates were analysed. Results: The fertilization rates were significantly different between OA and NOA patients (75.2% vs. 56.7%, p<0.05), however, embryo development did not differ between the groups (96.9% vs. 98.0%). Likewise, OA and NOA groups had no differences in their clinical pregnancy and delivery rates, 33.9% vs. 36.0% and 28.1% vs. 28.0%, respectively. With regard to sperm retrieved from fresh testicular tissue, fertilization rates were significantly different between the OA and NOA groups (76.4% vs. 52.9%, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. For sperm retrieved from thawed testicular tissue, the fertilization rates were significantly different between the two groups (74.7% OA group vs. 65.6% NOA group, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. Conclusions: Embryo development and clinical pregnancy did not differ in patients with obstructive and non-obstructive azoospermia, whether sperm retrieved from fresh and thawed testicular tissue were used, although the fertilization rates were different. Therefore, ICSI with sperm retrieved from fresh and thawed testicular tissue could achieve relevant clinical pregnancy results in patients with azoospermia. 목 적: 폐쇄성과 비폐쇄성 무정자증 환자에서 신선고환조직 정자와 융해고환조직 정자를 이용한 배발달과 임신결과를 비교 분석하였다. 연구방법: 폐쇄성과 비폐쇄성 무정자증 환자에서 총 222 주기의 TESE-ICSI를 시행하였다. 정자는 신선 또는 융해 고환조직으로부터 회수하였다. 수정은 ICSI 후 16~18시간째 확인하였으며, 배발달률과 임신율을 분석하였다. 결 과: 폐쇄성 무정자증군과 비폐쇄성 무정자증군에서 수정률은 유의하게 차이가 났으나 (75.2% 대 56.7%, p<0.05), 배발달률에서는 차이가 나지 않았다 (96.9% 대 98.0%). 마찬가지로 임상적 임신율 (33.9% 대 36.0%)과 분만율 (28.1% 대 28.0%)에서도 차이가 나지 않았다. 신선고환조직 정자의 경우, 수정률은 폐쇄성 무정자증군과 비폐쇄성 무정자증군에서 유의하게 차이가 났으나 (76.4% 대 52.9%, p<0.05), 배발달률, 임상적 임신율, 분만율에서는 차이가 나지 않았다. 융해고환조직 정자의 경우에서도 수정률은 폐쇄성 무정자증군과 비폐쇄성 무정자증군에서 유의하게 차이가 났으나 (74.7% 대 65.6%, p<0.05), 배발달률, 임상적 임신율, 분만율에서는 차이가 나지 않았다. 결 론: 폐쇄성과 비폐쇄성 무정자증 환자에서 신선고환조직 정자와 융해고환조직 정자를 이용하였을 경우 수정률에서는 차이가 있으나, 배발달과 임상적 임신 결과에는 차이를 보이지 않았다. 그러므로, 폐쇄성 또는 비폐쇄성 여부와는 관계없이 무정자증 환자에서 신선고환조직 정자 그리고 융해고환조직 정자를 이용하여 ICSI를 시행하면 적절한 임신 성적을 거둘 수 있다.

고환조직 동결-융해 후 회수된 고환 정자에 대한 Hypo-osmotic Swelling (HOS) Test의 효과

박용석,이형송,송상진,김정욱,강인수,서주태,Park, Yong-Seog,Lee, Hyoung-Song,Song, Sang-Jin,Kim, Jeong-Wook,Kang, Inn-Soo,Seo, Ju-Tae 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3

Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

Cell Soft System-3000과 Sperm Quality Analyzer-V를 이용한 정자 운동성 비교 분석

박용석,이선희,한상철,궁미경,김종우,서주태,Park, Yong-Seog,Lee, Sun-Hee,Han, Sang-Chul,Koong, Mi-Kyoung,Kim, Jong-Woo,Seo, Ju-Tae 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.2

Objective: To evaluate the results of CASA systems and to compare its results. Methods: Fifty semen sampales were analysed. Concentration, motility and forward progression were evaluated simultaneously on the same semen samples using Cell Soft System-3000 (CS system) and Sperm Quality Analyzer-V (SQA system). Results: Mean semen volume was $2.8{\pm}1.2\;ml$. Mean value of sperm concentration, motility, forward progression using CS system were $83.4{\pm}45.7{\times}10^6/ml$, $52.3{\pm}16.4%$ and $48.6{\pm}13.4%$, respectively. And mean value of sperm concentration, motility, forward progression using SQA system were $78.2{\pm}42.9{\times}10^6/ml$, $57.0{\pm}24.0%$ and $50.6{\pm}21.9%$, respectively. There were no statistical significancy of sperm concentration, motility, forward progression between the two devices. Conclusion: SQA system variables well correlated with the CS system. As a screening test for semen quality, CS system and SQA system is considered as useful in the management of male infertility.

동결-융해 후 회수된 고환 정자와 세정관내 정자의 수정 능력과 효율성에 관한 연구

박용석,전진현,이호준,강인수,김종현,이유식,서주태,Park, Yong-Seog,Jun, Jin-Hyun,Lee, Ho-Joon,Kang, Inn-Soo,Kim, Jong-Hyun,Lee, You-Sik,Seo, Ju-Tae 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.2

The combination of testicular sperm extraction (TESE) with ICSI can achieve normal fertilization and pregnancy rate and is effective method in obstructive and non-obstructive azoospermic patients. But, when pregnancy was not occurred, repeated testicular biopsies are not evitable. Therefore, in this study, we observed the survival rate of testicular spemratozoa and spermatozoa extracted from the seminiferous tubules after cryopreserved-thawed used for next IVF cycle with ICSI. In a total of 23 cases, obstructive azoospermia was 17 cases and non-obstructive azoospermia was 6 cases. In obstructive azoospermia, after thawing, motile spermatozua was observed in 13 cases (76.5%). The fertilization rate with 2PN was 67.6% and 5 pregnancies (29.4%) were achieved. In non-obstructive azoospermia, motile spermatozoa was observed in 2 case (33.3%) after thawing. The fertilization rates with 2PN was 53.7% and 3 pregnancies (50.0%) were achieved. A comparison of the results of motile spermatozoa after thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 3 cases (60.0%) and 12 cases (66.6%), respectively. The fertilization and pregnancy rates of thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 69.4% and 20.0%, 62.5% and 38.8%, respectively. Conclusively, thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section can achieve normal fertilization and pregnancy and cryopreservation of testicular spermatozoa and seminiferous tubule may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testis.

폐쇄성 무정자증 환자의 신선고환조직 정자와 동결고환조직 정자의 운동성이 임신율에 미치는 영향

박용석,이형송,변혜경,염혜원,송상진,임천규,이유식,윤종민,서주태,송지홍,강인수,궁미경,Park, Yong-Seog,Lee, Hyoung-Song,Byun, Hye-Kyung,Youm, Hye-Won,Song, Sang-Jin,Lim, Chun-Kyu,Lee, You-Sik,Yun, Jong-Min,Seo, Ju-Tae,Song, Ji-Hong,Kang, I 대한생식의학회 2001 Clinical and Experimental Reproductive Medicine Vol.28 No.2

Objective: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. Materials and Methods: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. Results: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). Conclusion: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospennic patients.

고환조직 정자채취술과 세포질내 정자주입술을 이용한 고환조직 정자의 수정률과 임신율

박용석,서주태,전진현,변혜경,김종현,이유식,손일표,강인수,이호준,Park, Yong-Seog,Seo, Ju-Tae,Jun, Jin-Hyun,Byun, Hye-Kyung,Kim, Jong-Hyun,Lee, You-Sik,Son, Il-Pyo,Kang, Inn-Soo,Lee, Ho-Joon 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.1

This study was carried to determine the possibility of finding motile spermatozoa and fertilization, pregnancy rate after testicular sperm extraction(TESE) with ICSI in obstructive and non-obstructive azoospermic patients. In 154 cases(132 patients), obstructive azoospermia was 77 cases and non-obstructive azoospermia was 77 cases. In obstructive azoospermia, patients generally showed normal spermatogenesis and included vas agenesis(n=8), multiple vas obstruction(n=7), epididymal obstruction (n=54). Total of 982 retrieved oocytes were obtained and 84.4% were injected. The fertilization rates with 2 PN and cleavage rate were 72.5% and 62.3%, respectively. 30 pregnancies(38.9%) were achieved and the ongoing pregnancies were 22 cases (28.6%). In non-obstructive azoospermia, patients showed hypospermatogenesis(n=49), maturation arrest(n=4), Sertoli cell only syndrome (n=24). The various stages of spermatogenic cell could be retrieved by TESE and could be reached normal fertilization and embryo development with ICSI. Total of 1072 retrieved oocytes obtained and 80.2% were injected. The fertilization rates with 2 PN and cleavage rate were 52.8% and 68.9%, respectively. 22 pregnancies(30.1%) were achieved and the ongoing pregnancies were 19 cases(26.0%). Conclusively, the combination of TESE with ICSI using testicular spermatozoa can achieve normal fertilization and pregnancy rate and effective method in obstructive and non-obstructive azoospermic patients.

효율 향상을 위한 BIPV용 마이크로 컨버터 실증에 관한 연구

박용석(Yong-Seog Park),박성배(Sung-Bae Park),김규호(Kyu-Ho Kim) 대한전기학회 2018 대한전기학회 학술대회 논문집 Vol.2018 No.-

폐쇄성 무정자증 환자와 정자형성저하증 환자의 고환정자를 이용한 체외수정 및 배아이식술의 결과

한상철,박용석,최수진,이선희,홍승범,이형송,임천규,송인옥,서주태,Han, Sang-Chul,Park, Yong-Seog,Choi, Su-Jin,Lee, Sun-Hee,Hong, Seung-Bum,Lee, Hyoung-Song,Lim, Chun-Kyu,Song, In-Ok,Seo, Ju-Tae 대한생식의학회 2009 Clinical and Experimental Reproductive Medicine Vol.36 No.1

목 적: 폐쇄성 무정자증 (obstructive azoospermia) 환자와 정자형성저하증 (hypospermatogenesis) 환자의 고환 정자를 이용한 체외수정 및 배아이식술의 결과를 비교, 분석하고자 하였다. 연구방법: 2003년 1월부터 2006년 12월까지 체외수정 및 배아이식술을 위해 고환조직 정자채취술을 시행한 폐쇄성 무정자증 환자 155명 (241주기)과 정자형성저하증 환자 28명 (34주기)을 대상으로 하였다. 고환조직 정자채취술과 세포질내 정자주입술을 실시한 후 수정률, 착상률, 임신율, 출산율을 비교하였으며, 통계적 분석은 t-test와 ${\chi}^2$-test를 사용하였다. 결 과: 정자형성저하증 환자의 고환정자 회수 결과, 신선고환을 사용한 21주기 중 1주기에서는 정자를 회수하지 못하였으나, 정자를 확인하고 고환조직 동결보존 후 융해한 13주기에서 모두 정자를 회수할 수 있었다. 수정률은 정자형성저하증 환자보다 폐쇄성 무정자증 환자에서 통계적으로 유의하게 높았다 (75.6 % vs. 62.6%, p<0.001). 난할 배아 발생률도 정자형성저하증 환자보다 폐쇄성 무정자증 환자에서 통계적으로 유의하게 높았다 (66.8% vs. 54.8% p<0.001). 그러나 우수배아 형성률, 임상적 임신율, 착상률, 출산율에는 차이가 없었다. 결 론: 정자형성저하증 환자의 고환정자를 사용하여 세포질내 정자주입술을 실시할 경우 수정률과 초기 난할율은 폐쇄성 무정자증 환자보다 낮지만, 정상적으로 발달한 배아를 이식할 경우 임신율, 착상률, 그리고 출산율에는 영향을 미치지 않는 것으로 사료된다. Objective: To compare the clinical outcomes of ICSI with sperm retrieved from testicular tissue in patients with obstructive azoospermia (OA) or hypospermatogenesis (HS). Methods: From January 2003 through December 2006, 155 patients with OA (241 cycles) and 28 patients with HS (34 cycles) were included in this study. We compared clinical outcomes of ICSI with testicular sperm such as fertilization rate, implantation rate, clinical pregnancy rate and delivery rate. Data were statistically analyzed using t-test and ${\chi}^2$-test. Results: Testicular spermatozoa could not be retrieved in 1 out of the 21 cycles where fresh testicular sperm extraction in HS patients. Fertilization rate (FR) was significantly higher in OA than HS (75.6 % vs. 62.6%, p<0.001). Cleavage rate (CR) per fertilized zygote was also significantly higher in OA than that in HS (66.8% vs. 54.8% p<0.001). However, there were no significant differences in good embryo rate (GER), clinical pregnancy rate (CPR), implantation rate (IR) and delivery rate (DR). Conclusion: Our results show that testicular sperm of HS does not affect CPR, IR, and DR although it has shown reduced FR and CR.

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.

동결보존이 생쥐 난소 조직 내 Heat Shock Protein 90의 발현에 미치는 영향

이선희,박용석,염혜원,송견지,한상철,배인하,Lee, Sun-Hee,Park, Yong-Seog,Yeum, Hye-Won,Song, Gyun-Jee,Han, Sang-Chul,Bae, In-Ha 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.1

Objective : Heat shock protein family is related to protective mechanism of cells by environmental changes. This study was performed to evaluate the effect of cryopreservation on the heat shock protein 90 (Hsp90) expression in mouse ovarian tissue. Methods : Cryopreservation of mouse ovarian tissue was carried out by slow freezing method. The mRNA level of Hsp90 expression in both fresh and cryopreserved mouse ovarian tissue was analyzed by RT-PCR. The protein expression of Hsp90 was evaluated by Western blot analysis and immunohistochemistry. Results: The mRNA and protein of Hsp90 were expressed in both fresh and cryopreserved mouse ovarian tissue. The amount of Hsp90 mRNA was increased in cryopreserved ovarian tissue after 60 and 90 minutes after thawing and incubation. The amount of Hsp90 protein was increased in the cryopreserved ovarian tissue after 6 hours of the incubation in Western blot analysis. In immunohistochemical study, Hsp90 protein was localized in cytoplasm of oocytes and granulosa cells. Significant level of immunoreactive Hsp90 protein was detected in theca cells contrast to the weak expression in ovarian epithelial cells. Conclusion: This results showed the increase of Hsp90 expression in both mRNA and protein level in the cryopreserved mouse ovarian tissue. It can be suggested that Hsp90 may play a role in the protective or recovery mechanism against the cell damage during cryopreservaion.