Identification of a High-yield Technique for Isolating EndometrialEpithelial Cells from the Mouse Uterus :A Comparison of Mechanical and Sedimentation-adherence Methods

손지온,조윤미,박혜진,안지연,송현진,임정묵,이승태 사단법인 한국동물생명공학회 2016 한국동물생명공학회지 Vol.31 No.1

An in vitro assay following culture of endometrial epithelial cells is essential for understanding epithelial cell function in reproduction. Several diverse techniques have been developed for isolating endometrial epithelial cells, although an optimal technique has not been identified. In this study, we describe a sedimentation-adherence (S-A) isolation technique with a high-yield cell-separating ability to isolate endometrial epithelial cells from 8-week-old female C57BL/6 mice. We analyzed total cell number, viability, morphology, and expression of cytokeratin 18 as an endometrial epithelial cell-specific marker in cells isolated using a mechanical method compared to the S-A technique. There were no significant differences in the total number, viability, or morphology of the putative endometrial epithelial cells with either method. In contrast, significantly more endometrial epithelial cells harvested using the S-A method were positively stained for cytokeratin 18 than those isolated using the mechanical method. These results confirm that the S-A method is more efficient for retrieving endometrial epithelial cells than a mechanical method.

심한 무력정자증 환자의 ICSI 시행시 Pentoxifylline을 사용한 정자처리법이 임상결과에 미치는 영향

손지온,신지수,정창진,조용선,엄기붕,최동희,김현주,Sohn, Jie-Ohn,Shin, Ji-Su,Jeong, Chang-Jin,Cho, Yong-Seon,Oum, Ki-Boong,Choi, Dong-Hee,Kim, Hyun-Joo 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.2

Objective: The aim of this study was to evaluate the effect of pentoxifylline (PF) on the conventional ICSI program undergone in severe asthenozoospermia. Method: Total 348 cycles of ICSI programs undertaken at CHA General Hospital from January, 1996 to September, 2000, were divided into two groups - injected with pentoxifylline-treated sperm (PFT, 204 cycles) or non-treated sperm (NPFT, 144 cycles) and the clinical results of PFT group were compared with those of NPFT. Results: PF-treatment on sperm increased their motility of normozoospermia and severe asthenozoospermia. Fertilization rate of PFT group was higher than those of ICSI programs undertaken using sperm of NPFT (70.6% vs. 62.9%, p<0.01). And, ET and clinical pregnancy rates of PFT were slightly higher than those of NPFT (93.1%, 44.2% vs. 90.3%, 36.2%). Conclusion: These results showed that treatment of pentoxifylline has a beneficial role on selection of viable sperm in severe asthenozoospermia.

동결-융해 배아 이식에서 손상된 할구의 미세 수술적 제거의 임상적 효과

최원윤,손지온,박은아,이동률,이우식,한세열,박이석,조정현,김수희,차광렬,윤태기,Choi, Won Yun,Sohn, Jie Ohn,Park, Eun A,Lee, Dong Ryul,Lee, Woo Sik,Han, Se Yul,Park, Lee Suk,Cho, Jung Hyun,Kim, Soo Hee,Cha, Kwang Yul,Yoon, Tae Ki 대한생식의학회 2005 Clinical and Experimental Reproductive Medicine Vol.32 No.1

Objective: Human infertility clinics have been faced the demand for improving clinical results. The purpose of this study was to evaluate the effect of microsurgical removal of damaged blastomeres (DB) in frozen-thawed embryos on the clinical outcomes. Methods: From January 2003 to May 2004, out of 258 thawing ET cycles were divided into three groups: Group-1 (n=46): Intact cleavaged embryos after thawing. Remained cycles with embryos containing DB were randomly divided into two groups. Group-2 (n=102): Drilling zona pellucida (ZP) of frozen-thawed embryos by acidified Tyrode's solution. Group-3 (n=110): Drilling ZP and removal of DB. Embryos after microsurgical manipulation were transferred into the uterus of patients. Results: Clinical profiles and the mean number of transferred embryos among three groups were not different. Pregnancy and implantation rates were similar in three groups. It were 30.4% and 9.3% in Group-1, 29.4% and 7.8% in Group-2, and 26.4% and 7.6% in group-3, respectively. Miscarriage rate in Group-3 (37.9%) was slightly higher than those in Group-1 and Group-2 (14.3% and 23.3%), but it was not statistically significant. Conclusion: Intact cleaving embryos after DB removal showed higher potent of pregnancy and implantation. We could not find any improvement of clinical outcome by removal of DB in frozen-thawed embryos.

난자 세포질내 정자주입술로 임신한 구형정자증 1예

남윤성,손지온,김현규,정미경,김승범,차광열 대한산부인과학회 2002 Obstetrics & Gynecology Science Vol.45 No.5

정자의 형태로 생식력을 판단하는 이론적인 근거는 정자가 투명대에 결합하고 통과하는데 필수적인 선단체의 능력을 간접적으로 반영하는 것이다. 기형정자는 투명대에 결합하거나 통과할 수 없다. 또한 정상형태의 정자가 투명대에 결합할 수는 있지만 통과하지 못하는 경우 수정이 되지 않는다. 정자두부의 가장 흔한 이상은 선단체와 관련된 경우이다. 정자두부의 이상 중 가장 잘 알려진 것은 구형정자증이다. 구형정자증에서는 선단체가 완전히 존재하지 않아서 두부는 원형이다. 그러므로 이런 경우 정자가 투명대를 통과할 수 없고 따라 서 불임이 되는 것은 당연하다. 그렇지만 구형정자증의 경우라 할지라도 난자 세포질내 정자주입술과 체외수정 시술을 통하여 임신이 가능하다.

고환 내 정자의 체외배양 중 운동성에 미치는 인간 난포액과 온도의 영향

신지수,손지온,이동률,김계성,정태규,김낙근,한지은,이우식,윤태기 韓國受精卵移植學會 2004 한국동물생명공학회지 Vol.19 No.1

The aim of this study was to evaluate the improvement of testicular sperm motility following different culture conditions such as human follicular fluid (hFF) and temperature. Testicular tissues obtained from azoospermia (n=21) were minced into small pieces by blade and recovered sperm suspension were cultured in Ham's F10 with or without 40% hFF at different temperatures (Group I: 37/with hFF, Group II: 32/withGroup III: 37/without, Group IV:32 /without The motility and viability of sperm were monitored during culture for 48 hours. Initial motility of testicular sperm was 10.91.9%. After 24 hours culture, sperm motility was 23.52.1% (Group I), 8.11.1% (Group II), 10.4 1.4% (Group III) and 4.00.8% (Group IV), respectively. After 48 hours, the motility had been changed as 322.3% (Group I), 14.31.7% (Group II), 5.3 1.4% (Group III) and 4.30.9% (Group IV). In hFF group (I and II), sperm motility of group I cultured at 37 was higher than those of group II at 32. But, sperm viability of group I cultured at 37 was lower than those of group II at 32 (54.44.1% vs. 59.43.7%) after cultured for 48 hours. We acquired the best motility of testicular sperm when performed in vitro culture for 48 hours in hFF supplemented medium at 37. Increase of sperm motility by in vitro culture could be useful tool fur human TESE-ICSI program.

ICSI를 실시한 2146예 중 수정란이식에 실패한 경우에 대한 원인 분석

이재호,윤태기,차광열,김현주,엄기붕,손지온 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.3

Since the introduction of intracytoplasmic sperm injection [ICSI] by Palermo et al.[1992], ICSI has been widely accepted in many IVF programs, including our own. Since our first successful pregnancy in 1993 using ICSI, we have continued to utilize the method in our IVF program to treat male factor patients. From January 1994 to December 1996, 2146 ICSI cycles were performed, most of the cases involving patients with severe male-factor infertility. The overall fertilization rate from the 2146 cycles was 76.0%. Embryo transfers failed in only 93 cycles[4.3%]. Details of the 93 failed ET`s were analysed. The fertilization rate of the ET failure group was 15.4% and the rates of 1PN and 3PN formation were 1.3% and 5.9%, respectively. A significant difference was found in the 3PN formation rate of the successful ET cycles [1.1%] compared with the 3PN rate of the failed embryo transfer group. The fertilization rate of small number[≤3] of oocyte group was slightly lower than the groups of oocyte number 4∼7 and more than 8. Forty eight cycles out of 93[51.6%] were categorized as having a poor quality oocyte with severe granulation, degenerative signs or abnormal morphology. In 24 cycles where non-motile sperm were used, fertilization did not occur. The women`s ages were not correlated with the fertilization rates. In conclusion, ET failures following ICSI for the treatment of severe male factor infertility was mainly caused by poor quality oocytes and/or poor viability of the spermatozoa used for injection into oocytes.

시험관아기 시술시 정자직접주입법을 이용한 2146 예의 임상 결과

최동희,윤태기,차광열,한세열,정형민,엄기붕,고정재,오종훈,손지온 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.1

Since Palermo and coworkers(1992) reported the first ICSI-baby, ICSI has been widely accepted as a treatment of male factor infertility. ICSI techniques have also been used to treat abnormal semen parameters in order to overcome fertilization failures and to increase clinical outcomes. From January 1994 to December 1996, 2146 ICSI cycles were performed at the Infertility Center of Cha General Hospital and we achieved a 75.1% of fertilization rate with 713 subsequent pregnancies(34.8%). The ICSI fertilization rate using 1-day old oocytes which failed to fertilize using conventional IVF method was lower than the ICSI fertilization rate using fresh oocytes(75.7% vs 65.9%). However, we achieved 13 pregnancies out of 125 embryo transfers from the total fertilization failure group. There was no statistical difference in the fertilization rates using normal semen or semen with single defect, but statistical differences were observed between normal semen and semen with two or more defects. No statistical difference was found in pregnancy rates between normal and defective semen groups. In our MESA, PESA and TESE programs, we achieved high fertilization rates(69.2∼82.5%) and a 45.8%(mean) pregnancy rate. Categorizing the female patients by age, ie., ≤29, 30∼34, 35∼39, ≥40, fertilization rates were consistently in the range of 74.5∼ 75.7%, but pregnancy rates significantly decreased in patients older than 35 yrs(31.3%) or 40 yrs(14.2%). Of the 713 pregnancies, we were able to obtain follow-up information of the babies in 527 cycles, of which we found 8abnormalities.