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X - 정자와 Y - 정자의 분리에 관한 연구 1 . Sephadex Gel 여과에 의한 인간정자의 분리
엄기붕,정길생,정병현 ( K . B . Oum,K . S . Chung,B . H . Chung ) 한국축산학회 1986 한국축산학회지 Vol.28 No.12
These experiments were carried out to develop new techniques useful for in vitro separation of X- and Y-bearing spermatozoa. One ㎖ of human semen was applied to the column (14 × 1.06 ㎝) filled with swellen Sephadex G-25 fine or G-50 fine gel and then elutriated with Locke solution. Elutriated solution was fractionated into 1 ㎖ by automatic fraction collector and spermatozoa included in each fraction were subjected to the estimation of viability and recovery rate and to F-body test. The results obtained in these experiments were summarized as follows: 1. Few spermatozoa were visible in the 1st to 3rd fractions and the highest concentration of spermatozoa was obtained from 7th to 9th fractions. 2. In case of Sephadex G-25 fine, the viable rate of spermatozoa before and after chromatography was 81.1 and 83.3%, respectively and in case of Sephadex G-50 tine, it was 79.9 and 82.35%, respectively. 3. The recovery rates of sperm applied to Sephadex G-25 and 50 fine were 40.8% and 52.3%, respectively. 4. The percentage of spermatozoa without F-body was increased from 53.0 to 67.1% and 52.6 to 68.2, respectively by the filtration through Sephadex G-25 and 50 column.
X- 정자와 Y- 정자의 분리에 관한 연구 2 . Percoll 중층원심분리법에 의한 인간정자의 분리
엄기붕(K . B . Oum),이주영(J . Y . Lee),고대환(D . H . Ko),정길생(K . S . Chung) 한국축산학회 1988 한국축산학회지 Vol.30 No.5
These experiments were carried out to develop new techniques for in vitro separation of X-and Y-bearing spermatozoa. One ㎖ of washed human sperm suspension was loaded on the isotonic discontinuous Percoll density gradient, and then it was centrifuged at 250×G for 25 min. After centrifugation, spermatozoa were fractionated according to Percoll density gradient. Spermatozoa included in each fraction were subjected to the estimation of motility, morphological abnormality, F-body test, and recovery rate of spermatozoa was also investigated. The results obtained in these experiments were summarized as follows: 1. Following centrifugation of discontinuous Percoll density gradient, population of spermatozoa increased progressively from low density to high density. The highest concentration of spermatozoa was observed in 7th fraction which included 20% of spermatozoa. 2. High percentage of motile spermatozoa was observed at high Percoll concentration and the highest percentage was obtained at 6th fraction. 3. Following Percoll centrifugation, percentage of X-sperm increased from 53.2% (control) to 74.1% (7th fraction). 4. Following centrifugation, sperm abnormality was increased at low Percoll gradient and decreased at high Percoll gradient. The lowest abnormality (15.0%) and the highest abnormality (46.0%) were observed at 7th fraction and 1st fraction, respectively.
시험관아기 시술시 정자직접주입법을 이용한 2146 예의 임상 결과
엄기붕(KB Oum),오종훈(JH Oh),손지온(JO Sohn),정형민(HM Jung),고정재(JJ Ko),한세열(SY Han),최동희(DH Choi),윤태기(TK Yoon),차광열(KY Cha) 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.
정자수정능력검사와 일반적인 체외수정과 정자직접주입법에서의 수정률과의 상관관계에 관한 연구
엄기붕(KB Oum),김현규(HG Kim),이재호(JH Lee),하정희(JH Ha),곽인평(IP Kwak),윤태기(TK Yoon),차광열(KY Cha) 대한산부인과학회 1998 Obstetrics & Gynecology Science Vol.41 No.3
We compared the results of three different sperm function test ie., sperm morphology test, hypoosmotic test, acridine orange test and the fertilization rates of IVF and ICSI to evaluate whether the sperm tests correlate with the fertilization rates of IVF and ICSI. In normal sperm morphology 0∼4%, the fertilization rates of IVF and ICSI were 5.6%[3/54] and 64.3% [27/42], respectively. In normal sperm morphology 5∼14%, the fertilization rates of IVF and ICSI were 43.0%[40/93] and 70.0%[49/70], respectively. There was a significant difference in the IVF fertilization rates in normal morphology 0∼4% and 5∼14% groups, however no difference was found in the ICSI fertilization rates according to the morphology. In HOS test, the IVF fertilization rates in low and high HOS value were 31.5% and 28.0%, respectively, and no difference was found in the fertilization rates when ICSI was employed according to the HOS value. However, if we compare the IVF and ICSI fertilization rates, significant differences were found. In high[≥ 60%] and low[<60%] AO value groups, the IVF fertilization rates were generally low[23.1 and 29.9, respectively], but higher fertilization rates were achieved in ICSI groups[69.9 and 67.4%, respectively]. No difference was found in the fertilization rates of IVF and ICSI according to AO value, but significant differences were found if we compare the IVF and ICSI fertilization rates. Our results showed that only sperm morphology scoring could predict the IVF fertilization rate, and no sperm characteristics could affect the ICSI fertilization rate.
정자직접주입법시술시 동결보존된 인간정소정자에 의한 수정 및 임신성공
엄기붕(Ki Boong Oum),윤태기(Tae Ki Yoon),차광열(Kwang Yul Cha),김현주(Hyun Joo Kim),남윤성(Yoon Sung Nam),김현규(Hyun Kyoo Kim),곽인평(In Pyung Kwak),한세열(Sei Yul Han) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.1
There are not much reports concerning with clinical results using frozen-thawed testivular sperm in ICSI program. It is speculated that the necessity of cryopreservation of testicular sperm to avoid repeating surgical procedure for obtaining sperm for ICSI. This study was carried out to confirm whether frozen-thawed testicular sperm could be fertilized and pregnancy could be achieved using embryos fertilized with frozen-thawed testicular sperm in ICSI program or not. Testicular sperm obtained from obstructive- or non-obstructive azoospermia patients were co-cultured for 3 days with Vero cells to improve sperm motility. By co-culturing with Vero cells for 3 days, O-ll% of sperm motility after thawing increased up to 8-42% after co-culturing. ICSI was performed using frozen-thawed, and co-cultured sperm with 66 oocytes obtained from 8 patients and 62 oocytes were survived and 49(79.0%) oocytes were fertilized normally. Embryo transfer was possible in 7 out of 8 patients, and pregnancy was achieved in 6 patients(85.7%). These results indicated that not only fresh testicular sperm but frozen-thawed testicular sperm can be used in ICSI program.
항정자항체가 일반적 체외수정 방법 및 정자직접 주입법(ICSI)에 미치는 영향에 관한 연구
오종훈,엄기붕,최동희,정미경,한세열,차광열,정길생,Oh, Jong-Hoon,Oum, Ki-Boong,Choi, Dong-Hee,Chung, Mi-Kyung,Han, Sei-Yul,Cha, Kwang-Yul,Chung, Kil-Saeng 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3
The purpose of this study was to examine the effects of anti-sperm antibody (ASA) on the fertilization processes using conventional IVF and ICSI procedure in human and hamster oocytes. In human IVF, we have observed restricted fertilization with sperm testing positive for ASA. ($23{\sim}90%$ IgA, 60-97 % IgG). However, if ICSI was perform in the next IVF cycle with the same patients, we could successfully fertilize the oocytes (37%; p<0.001), thus achieving pregnancy and delivery. When the sperm were cocultured in medium containing ASA, there were binding of ASA to sperm surface. In addition, the mean rate of the acrosomal reaction in an in vitro acrosome reaction test was lower for Ab-bound sperm (43.5%) than for Ab-free sperm group (51.3%, p<0.05). We used human sperm and hamster oocytes to confirm the negative effects of the ASA on fertilization. The sperm and/or oocytes have been expose to medium containing ASA before IVF and ICSI. In this experiment, the ASA was bound to the oocyte and sperm surface. The following results were obtain by using various combinations of ASA free or ASA bound sperm with ASA free or ASA bound oocytes for IVF. When ASA free sperm were inseminate with ASA free and ASA bound hamster oocytes, the fertilization rates are 89.6% and 74.3% respectively. However, when ASA bound human sperm were use the results were 62.5% and 55.6% respectively. These shows the fertilization rate was significantly decreased in both ASA bound and ASA free oocytes when using ASA bound sperm. No difference found when ASA are present on the oocyte surface. When the hamster oocytes was treated by ICSI with ASA free or ASA bound human spermatozoa, no significant difference was found. These results showed that ICSI is the most promising method for couples who fertilization was not possible by conventional IVF because of ASA.
Swim-up, Percoll, Sil-Select를 이용한 정자처리법에 의한 정자회수율, 운동성 및 체외수정율의 비교분석
하정희,엄기붕,정형민,정미경,김현규,고정재,윤태기,차광열,Ha, J.H.,Oum, K.B.,Chung, H.M.,Chung, M.K.,Kim, H.K.,Ko, J.J.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.2
It is well known that discard of seminal plasma from the semen and separation of motile sperm should be preceded before insemination for IUI or IVF. Till now, more than ten kinds of semen treatment methods have been developed. Of those, swim-up and Percoll methods have been used widely in ART laboratories as a routine semen treatment methods because of its advantages. However, there are reports that Percoll can make a genetic trouble because of its chemical structure and therefore the necessity has been arisen to substitute Percoll for other equivalent materials. This study was performed to evaluate the effects of three different sperm preparation methods (swim-up, Percoll and Sil-Select) on sperm motility, sperm recovery rate and fertilization rate. Also, the feasibility of using Sil-Select instead of Percoll in ART was evaluated. Each semen samples were divided into three fractions and motile sperm were recovered by swim-up, Percoll and Sil-Select gradient centrifugation methods. Normal and sub-normal criteria of fifteen semen samples and seventeen IVF cycles were included in these study. As results, no significant difference was found in sperm recovery rate in normal semen treated by a Swim-up, Percoll and Sil-Select method ($13.2{\times}10^6,\;17.5{\times}10^6\;and\;17.7{\times}10^6$ respectively). The initial sperm motility was 61.9% and this increased to 87.1%, 92.6% and 89.5% through Swim-up, Percoll and Sil-Select treatment, respectively. Higher motility was observed in Percoll and Sil-Select treated groups (81.5%, 79.2%, respectively) than swim-up group (66.8%) after incubation for 24hrs. In sub-normal group, sperm recovery rates were higher in Sil-Select group $(2.9{\times}10^6)$ than Percoll gradients group $(1.8{\times}10^6)$. In IVF cycles, the outcomes of fertilization using sperm treated by swim-up and Sil-Select group were similar (82.2%, 79.7% respectively). In conclusion, our results indicate that Sil-Select can be used as a substitute material for sperm preparation instead of Percoll.