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.

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.

전기사정과 시험관아기시술에 의한 성공적인 임신 1례

남윤성,김현주,전윤정,김현규,엄기붕,윤태기,차광열,Nam, Y.S.,Kim, H.J.,Jun, Y.J.,Kim, H.K.,Oum, K.B.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

Although anejaculation is a relatively uncommon occurrence in the general population, over 12,000 new cases are reported annually. Anejaculation may result from spinal cord injury, retroperitoneal lymph node dissection, diabetes mellitus, transverse myelitis, multiple sclerosis, or psychogenic disorders. At least 30% of men with this problem are or will be married and many will seek help to remedy their infertile state. The evolution of technique and instrumentation over the last 30 years has made electroejaculation an accessible and acceptable form of therapy. Recent successes in inducing ejaculation by means of rectal probe electrostimulation or vibratory stimulation combined with assisted reproductive techniques, such as zygote intrafallopian transfer (ZIFT), gamete intrafallopian transfer (GIFT), and in vitro fertilization (IVF), have provided these men means of producing their own biologic offspring. We have experienced a successful pregnancy with electroejaculation and in vitro fertilization in a infertile patient whose husband had an ejaculatory disturbance due to a spinal cord injury. So we report this case with a brief review of literatures.

인간 난관 상피세포와의 공동배양이 생쥐와 인간수정란의 체외발달에 미치는 영향에 관한 연구

고정재,정미경,도병록,엄기붕,윤태기,차광열,Ko, J.J.,Chung, M.K.,Do, B.R.,Oum, K.B.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1992 Clinical and Experimental Reproductive Medicine Vol.19 No.2

We examined effects of co-culture with human oviduct epithelial cells (HOEC) on the development of mouse and human embryos from early embryonic· stage to late morula or blastocyst stage (LM or B). In human, embryos were transferred and pregnancy rate was investigated. The HOEC, collected from surgically removed fallopian tube, were cultured in medium-199 supplemented with 20 % fetal cord serum (FCS). The HOEC were characterized by using immunocytochemical staining with anticytokeratin antibody and then used for cultures of mouse and human embryos. Results obtained from co-culture system were as follows. Development rate of mouse embryos was improved by co-culture system at late developmental stage (p<0.025). Human supernumerary embryos remained after transfer, unsuitable for freezing because of their poor quality, were co-cultured for 72hrs. Co-culture (78.79%) or conditioned medium (78.26%) system improved the developmemt rate, significantly, in comparision with control (11.11%)(p<0.00l). Co-cultured (85.71%) human zygotes for 24hrs showed the better development rate in comparision with control (50.00%) (p<0.01). When we transferred embryos cultured with the HOEC to patients, we obtained one pregnancy. Co-cultured human zygotes for 24hrs showed the better quality and viability for the replacement in comparision with control (p<0.01). In addition, improved pregnancy rate was obtained. Our results suggest that the co-culture system can rescue early degenerating embryos by improving early development and yield a resonable number of blastocyst for the appropriate replacement. The effect provided by cultured HOEC is not species specific for the development of embryos and it can be used to overcome in vitro blocks for the development. And also the co-culture system offers the possibility to freeze embryos at blastocyst stage which is more sucessful stage for the freezing. The HOEC monolayer may provide some stimulus via specific factor, which is unknown, to the development of embryos. Our results showed that the co-culture system with HOEC can be an alternative to conventional culture system.

폐색성 무정자증환자의 치료로써 미세적 부고환 정자흡입술(MESA) 및 정자직접주입법(ICSI)의 효과

김현주,김영찬,엄기붕,오종훈,이우식,한세열,최동희,윤태기,차광열,Kim, H.J.,Kim, Y.C.,Oum, K.B.,Oh, J.H.,Lee, W.S.,Han, S.Y.,Choi, D.H.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

To present and assess the efficacy of combination of microsurgical epididymal sperm aspiration(MESA) and intracytoplasmic sperm injection(ICSI) for the treatment of infertility due to unreconstructable obstructive azoospermia or congenital bilateral agenesis of vas deferens (CBAVD), MESA was performed in the 45 husbands ( 16 CBAVD, 29 unreconstructable genital tract obstruction), followed by ICSI of oocytes recovered from the wives hyperstimulated by GnRH agonist in combination with hMG and FSH. Cleaving embryos were transfered to the uterine cavity or follopian tube(ZIFT) 18 or 24 hours after ICSI procedure. In 45 cycles of MESA, 492 oocyte complexes were recovered. ICSI was carried out on 355 metaphase II oocytes and 226 oocytes (63.7%) showed normal two pronucleus fertilization. After 198 embryos were transferred in 43 cycles, an average of 5 per cycle, 20 patients presented a positive HCG and intrauterine pregnancy was confirmed by US. So, the clinical ongoing pregnancy rate per transfer was 46.5%. Until now, 8 patients have given birth to 9 babies, 5 male and 4 female, including 1 twin. The babies were all healthy except 1 twin female baby. There was 1 miscarriage at 7 weeks and chromosomal study of abortus revealed as 45X, monosomy. These results suggested that it was possible to achieve high normal fertilization and pregnancy rate by ICSI using epididymal sperm.

정자 직접 주입법 (ICSI) 이후에 수정에 실패한 인간 난자에 대한 염색체 분석

손원영,박성은,정형민,엄기붕,고정재,윤태기,차광열,Son, W.Y.,Park, S.E.,Chung, H.M.,Oum, K.B.,Ko, J.J.,Yoon, T.K.,Cha, K.Y. 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Despite the direct placement of sperm within the oocyte, fertilization failure still occurs after ICSI. This study was accomplished to analyze the chromosomes in oocytes failed to fertilize after ICSI comparing to oocytes failed to fertilize by conventional in vitro insemination. Seventy-four ICSI cycles and 122 conventional IVF cycles were included in analysis. Included unfertilized oocytes were from 74 patients (mean age = $32.7{\pm}3.7$). Ninety-three oocytes were informative and 83 oocytes were legible for cytogenetic analysis. Sixty-two oocytes out of 83 (74.7%) had normal chroruosomes, while 15 (18.1%) were hypoploidy, 6 (7.2%) were hyperploidy. Eighteen oocytes out of 93 (17.6%) were premature chromosome condensation (PCC). Two hundred ninety-four unfertilized oocytes after conventional insemination were subjected to chromosomal analysis and 180 oocytes were legible for analysis. One hundred thirty-two oocytes out of 180 (73.3%) were normal, while 22 (12.2%) were hypoploidy, 20 (11.1%) were hyperploidy, and 6 (3.3%) were polyploidy. Twenty-two oocytes (12.2%) were PCC. There was no difference in chromosomes between oocytes that failed to fertilize after ICSI or conventional insemination. High PCC rates in fertilization-failed oocytes suggest that oocytes maturity is another important factor in achieving successful fertilization.