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1. [구연] 관절제술 후 10년 이상 경과한 153예에서 정관복원술의 수술 경험

부산대학교 비뇨기과 박남철 외 4인 / 42차 불임학회 춘계학술대회 우수 논문 발표상 - 구연 1

정관절제술 후 10년 이상 경과한 153예에서 정관복원술의 수술 경험

부산대학교 의과대학 비뇨기과 / 박남철·엄박천·박현준·박언·이경미

목적:

정관절제술은 남성에서 수태조절의 목적으로 시행되는 영구불임술로서 1980년에서 1990년 초반 사이에 시행된 예에서 정관복원을 원하는 경우가 최근 점차 증가하고 있다. 따라서 정관절제술 후 10년 이상의 장기적 폐색 후 시행된 정관복원시 성공률과 이에 영향을 미치는 인자에 관한 연구가 필요한 실정이다.

대상 및 방법:

1983년 3월부터 2001년 3월까지 정관절제술 후 10년 이상 경과한 환자에서 정관복원술을 시행한 153예를 대상으로 의무기록과 전화 상담을 통해 해부학적 성공률 및 임신률 그리고 수술 성적에 영향을 미치는 인자를 조사 비교하였다.

결과:

대상군의 연령은 평균 41.6세 (30~57세), 정관절제기간은 평균 141.1개월 (120~240개월)이었다. 수술동기 (n=145)는 자녀를 더 원하는 경우가 63예 (43.5%)로 가장 많았고, 재혼이 36예 (24.8%), 아들을 원하는 경우가 27예 (18.6%), 자녀의 사망이 13예 (9.0%) 그리고 정관절제 후 합병증 6예 (4.1%) 순이었다. 정관절제기간 (n=153)은 10~14년 130예 (85.0%), 15~19년 15예 (9.8%), 20년 이상 8예 (5.2%)였다. 2001년 12월까지 추적조사가 가능하였던 98예에서 해부학적 성공률 및 임신률은 각각 81.6% (80/98) 및 36.0% (31/86)였다. 정관절제기간에 따른 각각의 해부학적 성공률 및 임신률은 10~14년군이 79.8% (67/84) 및 40.3 % (27/67), 15~19년군이 100% (8/8) 및 50% (3/6) 그리고 20년 이상군이 83.3% (3/6) 및 25% (1/4)이었다. 그 외 수술방법, 근위부 정관액 유무, 정자육아종의 유무, 정관문합 부위 및 봉합사 등의 수술 관련 인자에 따른 해부학적 성공률 및 임신률은 의미있는 차이를 나타내지 않았다 (p>0.05).

결론:

정관절제술 후 10년 이상 경과한 환자에서 정관복원술을 시행한 결과 해부학적 성공률 80%, 임신률 30% 이상의 비교적 우수한 수술성적을 나타내었다. 향후 최근의 보조생식술의 발달에도 불구하고 이들 환자들에 대해서 임신률을 개선시키기 위한 연구가 계속되어야 할 것으로 생각된다.

2. [구연]Aberrant Apoptosis in Chorionic Villi is Involved in Recurrent Miscarriage

포천중문의대 차병원, 광주 시엘병원 김정욱 박사 외 / 42차 불임학회 춘계학술대회 우수 논문 발표상 - 구연 9

Aberrant Apoptosis in Chorionic Villi is Involved in Recurrent Miscarriage

1Kim JW, 1Lee J, 1Choi BC, 2,3Baek KH, 2,3Choi HK, 2,3Lee SH, 2,3Ko JJ, 2,3Cha KY
1Center for Recurrent Miscarriage and Infertility, Creation and Love Womens Hospital, Gwangju, Korea, 2College of Medicine, Pochon CHA University, 3Infertility of Medical Center, CHA General Hospital, Seoul, Korea

Introduction:

Apoptosis, programmed cell death, is a cellular process in development and tissue home- ostasis. Abnormalities in cell death control can lead to various diseases, including cancer, autoimmunity, and degenerative disorders. Apoptosis is a rare event in several reproductive tissues including human placenta. The causes of unexplained recurrent miscarriage is not fully understood. However, most of rese- arches have been focused on immune mechanisms of maternal-fetal interface. In these studies, we in- vestigated whether apoptosis in chorionic villi is associated with recurrent pregnancy loss.

Materials and Methods:

We previously performed cDNA subtractive hybridization analysis in fetal chorionic villi from normal patients and RPL patients, showing different expression level of apoptosisrelated genes. Therefore, we investigated whether apoptosis is aberrant in chorionic villi from recurrent pregnancy loss (RPL) patients. Expression level of genes including Fas, FasL, Bax, Bid, Bad, caspase 3, caspase 6, caspase 7, caspase 8, caspase 9, caspase 10, and caspase 12 was analyzed by the intensity of the EtBr staining by Gel-Doc and reverse northern blot analysis.

Results:

In these results, we demonstrated the different expression level of 12 apoptosis-related genes. Expression levels for apoptosis-related genes showed higher in chorionic villi from RPL patient than those from normal control. In addition, expression pattern by reverse northern blot analysis will be analyzed.

Conclusions:

RT-PCR analysis revealed that apoptosis-related genes expressed more in the chorionic villi with RPL patients than normal controls, suggesting that aberrant apoptosis could be one of reasons that may lead to recurrent spontaneous abortion during early human development. The molecular mechanisms for this phenomenon have to be elucidated for finding their therapeutic methods.

3. [포스터] Maintenance of Human Embryonic Stem Cells Derived from Frozen-thawed Blastocysts on Feeder-free Culture Condition

마리아병원, 건국대학교 김은영 외 / •42차 불임학회 춘계학술대회 우수 논문 발표상 - 포스터 11

Maintenance of Human Embryonic Stem Cells Derived from Frozen-thawed Blastocysts on Feeder-free Culture Condition

마리아 기초의학연구소/마리아 생명공학연구소, 1건국대학교, 2마리아 병원
김은영·이금실·박은미·신현아·민현정·박세필·정길생1·임진호2

Objective:

This study was to confirm whether the established human embryonic stem (hES) cell growth can be maintained without mouse embryonic feeder (MEF) cells.

Materials and Methods:

The hES cells (MB02 and MB03) derived from frozen-thawed blastocysts were subcultured until 10th passage (about 2 month, 40 population doublings) on MEF feeder. And then some of these hES colonies were cultured on feeder-free condition using Matrigel-coated plate/STO cell condi- tioned medium. Characterization of hES cells cultured on feeder or off feeder were taken by alkaline pho- sphatase staining, karyotyping, cell surface marker staining, Oct4 gene expression and telomerase activity.

Results:

The hES cells cultured on feeder-free condition during subculture (about 6 month, 120 popula- tion doublings) indicated stable proliferation rate, normal karyotype. high telomerase activity. Similar to cells cultured on feeders, hES cells maintained under feeder-free conditions expressed Oct4, alkaline pho- sphatase, surface marker (SSEA-4, TRA-1-60, TRA-1-81). Also, embryoid bodies cultured on gelatin dish in differentiation medium were examined by RT-PCR, some specific factors represented three embryonic germ layers were determined in vitro (NF-M, keratin, enolase, cAct and amylase).

Conclusion:

The established hES cells derived from frozen-thawed blastocysts can be maintained on feeder-free condition without loss of human cell characteristics.

4. [포스터] Effectsof Growth Factor o the Spermatogenic Cells from Infertile Men in co-culture System with TM4 Monolayer

삼성제일병원 / •42차 불임학회 춘계학술대회 우수 논문 발표상 - 포스터 18

Effects of Growth Factor on the Spermatogenic Cells from Infertile Men in co-culture System with TM4 Monolayer

Song SJ1, Min DM1, Park YS1, Kang IS2, Lee YS3, Seo JT3
1Laboratory of Reproductive Biology & Infertility, 2Department of Obstetrics & Gynecology, 3Department of Urology, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine, Seoul, Korea

Objectives:

The development of assited reproduction technique significantly improved pregnancy rate in male infertility. However, the problem of severe idiopathic male infertility were not solved inspite of many efforts. In vitro culture of spermatogenic cells were then initiated in an effort to try to overcome the low clinical outcome. Recently, the production of flagellar growing spermatid by in vitro culture system was achieved. Also, auto/xenotransplantation technique of spermatogonial cell were developed. The esta- blishment of spermatogenesis in vitro is very important for clinical outcome and understanding of molcular events in male reproductive organ. Therefore, we evaluated the effect of growth factors (Stem Cell Factor (SCF), Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF)) in spermatogenic cell co-culture system and estimate for proliferative abilty of encasulation culture system with Sertoli cell monolayer.

Materials & Methods:

From Jan. until Oct. 2001, 19 obstructive azoospermic patients and 4 nonobstructive azoospermic patients entering IVF/ICSI programme were enrolled. Spermatogenic cells were co-cultured with or without empty zona pellucidae on TM4 Sertoli cell monolayer supplemented with different concentrations of SCF and GM-CSF up to 168 hrs. Different survival rates were compared by trypan blue exclusion test and Hoechst staining. In order to compare the proliferation and differentiation abilty in different conditions, immunocytochemical staining with anti-c-kit antibody was performed. The presence of early spermatogenic cell was confirmed by RT-PCR with c-kit primer.

Results:

The survival rate in encapsulated system attenuated the time-dependent death rate than free culture system. The survival rate of spermatogenic cells from obstructive azoospermic patients were sligh- tly higher than that of non-obstructive azoospermic patients at 168 hrs culture. Addition of growth factor in encapsulated co-culture system increased proliferation rates of spermatogenic cells. In this system, the most optimal concentrations of SCF, GM-CSF were 1 ng/ml, 1 ng/ml, respectly. Presence of early sperma- togenic cells was confirmed by expression of c-kit transcript during all culture period.

Conclusions:

The encapsulation of spermatogenic cells with zona pellucidae in co-culture system enhanced cell survival and proliferation rates probably by protecting from loss of germ cells and various damage during in vitro culture. The addition of growth factor in appropriate concentrations improved cell growth as well.