생쥐 초기배아와 사람의 수정란의 발생에 미치는 생식수관 상피세포의 영향에 관한 연구

이호준,변혜경,김정욱,황정혜,전종영,김문규,Lee, H.J.,Byun, H.K.,Kim, J.W.,Hwang, J.H.,Jun, J.Y.,Kim, M.K. 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.3

Mammalian oviductal epithelial cells have been known to improve in vitro fertilization and embryonic development. Recently, co-cultured human embryos with the epithelial cells in human genital tract has been reported to improve the pregnancy rate. The purpose of the study was to investigate the effects of the epithelial cells of human genital tract on the development of mouse early embryos and human fertilized oocytes. The epithelial cells of human genital tract were collected from the fallopian tubes which were obtained during hysterectomy in fertile women and from the endometrium during endometrium biopsy. Collected human ampullary cells(HACs) and endometrial cells(HECs) were cultured for 10 days to establish primary monolayer. Second passaged HACs and HECs were obtained by trypsinization were cryopreserved in PBS with 1.5 M DMSO for later use. To investigate the effect when co-cultured with HACs and HECs, we tried to apply strict quality control on mouse embryo, from two cell to blastocyst prior to human trial. The results of quality control were as follows; In Group I (Ham's F10 with 10% FCS), Group IT (co-cultured with HACs) and Group ill (co-cultured with HECs), developmental rates to blastocyst were 63.3%(253/400), 76.0%(304/ 400),74.0%(296/400), respectively. Hatching rates were 36.8%(147/400), 41.80/0(167/400), 38.0%(152/400), respectively(p<0.05). To perform the human IVF, cryopreserved HACs were thawed at 37$^{\circ}C$ waterbath, seeded on the well dish and cultured for 48 hI'S. The pronuclear stage embryos were transferred to the seeded well dish. After 24 hRS, co-cultured embryos were examined and transferred to patient's uterus. The results of human IVF when co-cultured with HACs were that fertilization and developmental rates were 61.8% (256/414), 95.3% (244/256) as compared with 57.2% (279/488) and 94.6%(264/279) in Ham's F10 supplemented with 10% FCS(control). However, 62.9% (161/256) of co-cultured human embryos showed good embryos(no or slight fragmentation) as compared with 53.8 % (150/279) in control(p < 0.05). Pregnancy rate was 40.0% (12/30) when co-cultured with HACs whereas 30.6%(11/36) in control. In conclusions, co-culture system using HACs and HECs improved the developmental and hatching rates of mouse embryo. Also, in human IVF system when co-cultured with HACs, it improved both the quality of human embryos and the pregnancy rate.

인간의 체외수정배아이식술에서 보조부화술이 임신률에 미치는 영향에 관한 연구

이호준,김정욱,변혜경,전진현,손일표,전종영,Lee, H.J.,Kim, J.W.,Byun, H.K.,Jun, J.H.,Son, I.P.,Jun, J.Y. 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

In human IVF-ET, the development and morphology of the embryo have been known to affect implantation and pregnancy rates(PRs). Recently, pregnancy has been reported to related to the embryos with thick zona-pellucida, high levels of fragmentation, poor blastomere development and zona hardening. Although the mechanism of implantation is unclear, it is thought that the hatching process precedes implantation and that the hatching is related to implantation and PRs. This study was carried out to investigate the effect of assisted hatching(AHA) on the improvement of PRs in human IVF-ET. The results were as follows; 1. The PRs of the AHA group (40.8%) was significantly higher than that of control group(27.2%)(P<0.01). 2. According to the age of patients, the PRs of control and AHA groups were 33.9%(20/59), 44,4%(12/27) in <30 yrs, 26.1%(30/115), 38.3%(18/47) in 31-35 yrs, 22.4%(13/58), 41.4%(12/29) in >36 yrs, respectively. 3. According to the factors of infertility in AHA group, unexplained(immunologic factor) (40.0%) and male factors(41.9%) were higher than female(tubal obstruction, endometriosis, adhesion) factor (28.9%). As a result, it is suggested that AHA technique improve the PRs in poor prognosis patients. It is concluded that AHA method can be used to improve the PRs in human lVF-ET.

폐쇄성 정로장애로 인한 무정자증 환자에서 미세수술적 부고환 정자흡입술과 세포질내 정자주입술을 이용한 수정율 및 임신율 증진에 관한 연구

손일표,홍재엽,이유식,전진현,박용석,이호준,강인수,전종영,Son, I.P.,Hong, J.Y.,Lee, Y.S.,Jun, J.H.,Park, Y.S.,Lee, H.J.,Kang, I.S.,Jun, J.Y. 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.3

We studied the role of assisted fertilization(subzonal insemination, intracytoplasmic sperm injection) in enhancing fertilization and pregnancy rate in obstructive azoospermia. MESA was performed in the patients with congenital absence of the vas deferens and unreconstructable obstructive azoospermia. Sperm were aspirated microsurgically from various sites along the epididymal stump. Sperm were then washed on a mini-PercoH gradient or swim-up method and treated by 2-deoxyadenosine and pentoxifylline. Conventional IVF(group I, 14 cycles), SUZI(group II, 13 cycles) and ICSI(gruop III, 28 cycles) were carried out in 55 treatment cycles. The clinical results are as follows: 1. Fertilization rates for group I, II and III were 16.1 %,31.4% and 48.6%, retrospectively (p<0.05). 2. Clinical pregnancy rates for group I, II and III were 7.1 %,7.7%, and 32.1 'Yo, retrospectively. 3. In 5 of MESA-ICSI cycles, epididymal sperm from alloplastic spermatocele were used and 2 clinical pregnancies (40%) were obtained. According to our results the combined MESA-ICSI procedure is highly effcient in improving fertilization and pregnancy rate in congenital absence of the vas deferens and unreconstructable obstructive azoospermia.

자궁내막증 또는 자궁선근증에 의한 불임환자에서 장기간 GnRH analogue ( Long - term Down DRegulation ) 사용 후 과배란 및 체외 수정 시술의 결과

양광문(K . M . Yang),유근재(K . J . Yoo),최범채(B . C . Choi),김계현(K . H. Kim),이홍복(H . B . Lee),이재훈(J . H . Lee),송인옥(I . O . Song),송지홍(J . H . Song),궁미경(M . K . Koong),전종영(J . Y . Jun),강인수(I . S . Kang) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.10

목적 : 자궁내막증 또는 자궁선근증을 가진 불임 환자의 치료에 있어 GnRH analogue를 사용한 LTDR의 과배란 유도시 그 효과를 알아보았다. 방법 및 재료 : 복강경과 질식 초음파를 이용하여 자궁내막증 또는 자궁선근증을 진단 받은 불임 여성 43명의 47 LTDR cycles이 연구 대상이 되었다. GnRH analogue를 3개월 이상 장기간 사용하여 down regulation 시킨 후 gonadotropin을 이용하여 과배란 유도를 시행한 경우를 LTDR로 정의하였으며 임신을 위해 자궁강 내 정자 주입술 또는 체외 수정 및 배아 이식이 시행되었다. 결과 : 환자의 평균 연령(meanSD)은 33.83.8이었고 자궁선근증이 10cycles였으며 자궁내막증은 34cycles(1기:12cycles, 2기:9cycles, 3기 :5cycles, 4기:8cycles)이었다. 체외 수정 시 이식된 배아는 평균(meanSD) 3.70.3개였다. 자궁내막증을 가진 34cycles중 17.6%(6/34), 자궁선근증을 가진 10cycles중 40.0%(4/10)의 임신율을 보였다.결론 : 이 연구에서 LTDR은 자궁내막증과 자궁선근증에 의한 불임 환자의 치료에 효과적인 것으로 보이나 연구 대상군의 확대 조사가 필요하리라 사료된다. Objective : To assess the clinical efficacy of long term down regulation (LTDR) for in vitro fertilization (IVF) in infertile patients with endometriosis or adenomyosis.Materials and methods : Analysis were made from data collected from 45 paients who had endometriosis or adenomyosis diagnosed using pelvic laparoscopy and ultrasonography and had undergone intrauterine insemination or in vitro fertilization. LTDR was defined as long term, as long as three months down regulation of ovarian function followed by induction of ovulation using gonadotropins. Of these patients, 43 had undergone LTDR followed by IVF-ET, whereas 30patients in control group had undergone short or long protocol as an ovarian stimulation regimen.Results : The multiple independent parameters such as plasma basal estradiol (E2), plasma E2 level on day of hCG administration, number of oocytes retrived, number of good quality oocytes, number of embryo and number of transfered embryos were not significantly different between LTDR treated group and other control group. The clinical pregnancy rate in women treated with LTDR (17.6% in endometriosis, 40% in adenomysis, and 33.3% in endometriosis combined with adenomyosis) were comparable with those of control group (15.4% in endometriosis, 33.3% in adenomyosis and 40% in endometriosis combined with adenomyosis). Conclusions : In this study the LTDR as a ovarian stimulation regimen for IVF does not seems efficienct in infertile patients having endometriosis or adenomyosis than other protocols such as short or long, however further study with large number may be needed.

Toxic and Non-Toxic Peritoneal Fluid:Effects on Sperm Motility

노성일,최규완,박종민,이승재,전종영,Roh, S.I.,Choi, K.W.,Park, J.M.,Lee, S.J.,Jun, J.Y. The Korean Society for Reproductive Medicine 1989 Clinical and Experimental Reproductive Medicine Vol.16 No.2

불염여성에서 복강내액의 독성 유무를 알기 위해서 콤퓨터 정액 분석기를 이용하여 복강내액의 정자 활동성을 분석하였다. 연구대상은 자궁내막증 환자 14예와 자궁내막증이 없는 불염환자로부터의 복강내액 9예로 하였다. 연구방법은 건강인의 정자를 셰척 분리하여 대상환자의 복강내액과 Hams F10 배양액을 동량 흔합하여 (50%) 배양후 정자 활동성을 측정하였다. 정자 활동성의 감소는 배양전 0시간의 수치와 비교해서 1시간, 4시간 그리고 24시간후의 결과를 비교 분석하였다. 자궁내막증 제 1기 및 제 2기 환자와 자궁내막증이 없는 환자의 복수에서의 정자 활동성의 감소는 통계학적으로 의의있는 차이는 없었다. 그러나 자궁내막증이 심한 제 3기의 환자군에서는 의의있는 차이가 있었다(p<.05). 실험군간의 정자 활동성의 평균치 차이는 없었으나,각 군에서의 환자별 개인 성적은 정자 활동성이 현저히 감소한 독성있는 복강내액과 비독성의 복강내액을 쉽게 구별할 수 있었다. 결론적으로 정자 배양을 이용한 복강내액 독성 검사는 비교적 간단하고 경제적이므로 염상적으로 최근에 대두된 복강내 수정 치료법의 예비검사로서 유용할 것으로 사료된다.