The Effects of Isotypes and Regional Distribution of Antisperm Antibodies on Semen Parameters and Fertilizing Ability

방명걸,문신용,Pang, Myung-Geol,Moon, Shin-Yong The Korean Society for Reproductive Medicine 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1

항정자항체의 종류 및 존재부위가 정액성상 및 수정능력에 미치는 영향을 조사하였다. 항정자항체의 종류 및 존재부위는 immunobead binding test에 의하여 시행하였으며, 정자와 수정능력은 투명대제거 햄스터 난자 침입법에 의하여 시행하였다. 항정자항체는 정자수, 운동성 및 운동지수에 악영향을 끼쳤으며, 수정능력에도 악영향을 끼쳤다. 항정자항체의 존재부위에 따른 차이는 보이지 않았다. 항정자항체 IgG가 정자두부 혹은 정자미부에 존재할 경우 및 항정자항체 IgA가 정자미부에 존재할 경우 수정능력을 크게 감소시켰다. To investigate the influences on semen parameters and fertilizing capacity of immunoglobulin (Ig) isotypes and regional distribution of antisperm antibody (ASA) on the human sperm surface. Sixty-seven ASA-positive patients were compared with 96 ASA-negative donors. ASAs in semen showed significant negative effects on both semen parameters and fertilizing capacity; in those with ASAs in the sperm head and/or tail, the reductions were significant. In the head as well as the tail, there was close correlation between fertilizing capacity and both IgG and IgA. Both semen parameters and fertilizing capacity are significantly affected by the presence of ASA in semen. In particular, antibodies IgG to sperm head and/or tail, and antibodies IgA to sperm tail appeared to have a highly detrimental effect on fertilizing capacity.

Sperm Penetration Assay의 임상적 타당성에 관한 연구

방명걸,오선경,신창재,김정구,문신용,장윤석,이진용,Pang, Myung-Geol,Oh, Sun-Kyung,Shin, Chang-Jae,Kim, Jung-Gu,Moon, Shin-Yong,Chang, Yoon-Seok,Lee, Jin-Yong 대한생식의학회 1993 Clinical and Experimental Reproductive Medicine Vol.20 No.1

The present study was designed to test the validity of the semen analysis(S/A) and the sperm penetration assay(SPA) as a prognostic indicator of male fertility in 123 patients undergoing in vitro fertilization(IVF). We attempted to correlate the traditional semen parameters or the extent of sperm penetration in SPA with the results of human IVF rate or cleavage rate. Poor correlation was found between the results of S/A and human IVF rate(sensitivity, 80.6% ;specificity, 46.7%; positive predictive value, 91.6%;negative predictive value, 25%). Conversely, good correlation was found between the results of SPA and human IVF rate(sensitivity, 100% ; specificity, 80% ;positive predictive value, 97.3% ;negative predictive value, 100%). Our results corroborate the conclusion that SPA can be a valuable tool as a prognostic indicator of male fertilizing ability.

TEST-Yolk Buffer에 의한 인간 정자의 수정능 증진효과에 관한 연구

방명걸,김기철,신창재,문신용,이진용,장윤석,Pang, Myung-Geol,Kim, Ki-Chul,Shin, Chang-Jae,Moon, Shin-Yong,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1992 Clinical and Experimental Reproductive Medicine Vol.19 No.1

The present study was undertaken to clarify the role of TEST-Yolk Buffer(TYB) as a factor for the improvement of human sperm fertility potential. We examined the effects of low temperature capacitation using TYB on sperm motility (%), motility pattern, normal morphology, true acrosome reaction, sperm penetration assay and human in vitro fertilization. Comparing the TYB method and swim-up method, the sperm motility(%) of selected sperm was not significantly different, but statistically significant differences were found in curvilinear velocity, linearity, lateral head displacement, normal morphology(%) and true acrosome reaction(%)(p<0.05). Results obtained from the sperm penetration assay demonstrated that the penetration index and penetration rate were increased significantly(p<0.05) when the spermatozoa were incubated in TYB, as compared with swim-up method. And fertilization of intact human oocytes was more succesful when spermatozoa were pretreated with TYB at $4^{\circ}C$ for 48 hours as compared with swim-up method. Our results show that TYB method have advantages in terms of enhancement of sperm hyperactivation, increased true acrosome reaction, increased ability to penetrate zona-free hamster ova and augmented fertilization of human oocytes, suggesting that TYB is superior in its ability to preserve sperm motility and fertilizing ability.

냉동보존된 햄스터 난자를 이용한 인간정자의 생식력 평가

방명걸,정구민,김석현,신창재,김정구,문신용,이진용,장윤석,Pang, Myung-Geol,Chung, Ku-Min,Kim, Seok-Hyun,Shin, Chang-Jae,Kim, Jung-Gu,Moon, Shin-Yong,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1992 Clinical and Experimental Reproductive Medicine Vol.19 No.2

To solve the logistical problems of the sperm penetration assay (SPA) to provide just a sufficient number of hamster ova exactly when they are needed, a new method to cryopreserve the ova has been devised (1-step dehydration and 2-step thawing). After freezing & thawing of zona-intact (ZI) and zona-free (ZF) hamster ova according to this new method, the frozen-thawed ova were compared with fresh, control ova (FO) in terms of the degree of sperm penetration in SPA using semen samples from fertile donors, subfertile, and infertile male. Each sperm sample was capacitated for 42 hours inTEST-Yolk Buffer before insemination in SPA. In fertile doner, both the penetration rate and penetration index were lower in SPA using frozen ova (ZI; 92.4%, 6.2, ZF; 63.7%, 3.9) than those of SPA using fresh ova (99.3%, 8.4). There was a significant correlation between the penetration index of SPA using FO and ZI (p<0.001), and between those of SPA using FO and ZF and ova (p<0.001). In subfertile patient, both the penetration rate and penetration index were lowered in frozen ova (ZI; 62.3%, 1.3, ZF; 21.8%, 0.4) than those of fresh ova (74.8%, 1.8). There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05 and p<0.001, respectively). In infertile patient, both the penetration rate and penetration index were ZI; 3.1%, 0.0, ZF;0.0%, 0.0, respectively. There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05).

Fluorescence in Situ Hybridization 시행을 위한 인간정자 탈응축법의 적정화

방명걸,Pang, Myung-Geol 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West et al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS). Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at $37^{\circ}C$ for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at $4^{\circ}C$. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious "nullisomy" due to hybridization failure without inducing spurious "disomy" resulting from increased distances between split signals.

면역학적 불임환자에서 체외수정 시술에 의한 임신 1예

방명걸,오선경,김석현,신창재,김정구,문신용,이진용,장윤석,Pang, Myung-Geol,Oh, Sun-Kyung,Kim, Seok-Hyun,Shin, Chang-Jae,Kim, Jung-Gu,Moon, Shin-Yong,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1992 Clinical and Experimental Reproductive Medicine Vol.19 No.2

In vitro fertilization and embryo transfer was performed in a patient with immunologic infertility. This patient delivered at preterm a normal healthy male infant.

인간정자에 있어서 정자처리법의 비교

방명걸,정구민,신창재,김정구,문신용,장윤석,이진용,이상훈,정영채,김창근,Pang, Myung-Geol,Chung, Ku-Min,Shin, Chang-Jae,Kim, Jung-Gu,Moon, Shin-Yong,Chang, Yoon-Seok,Lee, Jin-Yong,Lee, Sang-Hoon,Chung, Yung-Chai,Kim, Chang-Keun 대한생식의학회 1993 Clinical and Experimental Reproductive Medicine Vol.20 No.2

Procedures to separate motile. normal & motile and acrosome-reacted sperm with high efficiency have clinical application in Assisted Reproductive Technology in terms of increasing the probability of fertilization by a normal sperm and subsequent normal embryonic development. This study evaluated the effects of 10 sperm preparation techniques [Swim-up from a washed pellet (SU). Swim-up from semen (SO). Continuous Percoll Gradients I (PIC). Discontinuous Percoll Gradients I (PID). Continuous Percoll Gradients II(P II C). Discontinuous Percoll Gradients II(P II D), SpermPrep (SFC). Wang's tube (WT). Albumin Gradients (AG), Low temperature capacitation (LTC)] on motility (%), normal morphology (%), motile sperm recovery rate(%). morphologically normal & motile sperm recovery rate (%), true acrosome reaction (%) and fertilizing ability. A P II D proved to be an effective means of separating morphologically normal & motile sperm. Our results indicated the P II D has advantages as compared with other methods in terms of recovery rate. enhancement of motility and normal morphology. And a LTC seems to be an effective means of enhancing the true acrosome reaction and fertilizing ability. These results suggest that the combined method of LTC and P II D for separation of morphologically normal & motile sperm and acrosome reacted sperm may be a useful procedure for intrauterine insemination and in vitro fertilization in the management of male factor infertility as well as for isolation of subpopulation of sperm for basic research.

체외수정시술에서 정자의 수정능력이 배아의 발생능 및 임신율에 미치는 영향

방명걸,정병준,문신용,Pang, Myung-Geol,Jung, Byeong-Jun,Moon, Shin-Yong 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.1

Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

국산 Fluorescence in Situ Hybridization 시스템을 이용한 다양한 검체에서의 염색체 분석

문신용,방명걸,오선경,류범용,황도영,정병준,최진,손철,장준근,김종원,김석현,최영민,Moon, Shin-Yong,Pang, Myung-Geol,Oh, Sun-Kyung,Ryu, Buom-Yong,Hwang, Do-Yeong,Jung, Byeong-Jun,Choe, Jin,Sohn, Cherl,Chang, Jun-Keun,Kim, Jong-Won,Kim, Se 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Z1, DXZ1, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.

한국인 남성을 대상으로 한 햄스터 난자 침투 분석법의 정상 가임역 설정

김석현,방명걸,신창재,김정구,문신용,이진용,장윤석,Kim, Seok-Hyun,Pang, Myung-Geol,Shin, Chang-Jae,Kim, Jung-Gu,Moon, Shin-Yong,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.1

To establish the normal fertile range in the results of the sperm zona-free hamster ova penetration assay (SPA) in Korean male, SPA using the low temperature ($4^{\circ}C$) capacitation in TEST-yolk buffer (TYB) was performed in 67 fertile and 26 infertile men. Sperm parameters in routine semen analysis were also checked and compared with the results of SPA. Sperm concentration, motility and motility index (MI) were significantly higher in fertile group compared with infertile group: $96.0{\pm}46.6$ vs $43.6{\pm}31.9{\times}10^6/ml$, $65.5{\pm}14.8%$ vs $45.8{\pm}23.6%$ and $46.31{\pm}13.29$ vs 27.40{\pm}17.98$, respectively. In fertile group, the hamster ova penetration rate (PR) was $98.5{\pm}5.0%$ (80%-100%), and the penetration index (mean penetrations per ovum, PI) was $9.59{\pm}6.35$(3.1-29.0). All the fertile men showed PI>3.0. In infertile group, PR was $24.6{\pm}24.8%$ (0%-70%), and PI was $0.40{\pm}0.42$ (0-1.3). Both PR and PI were significantly lower in infertile group. There was a significant correlation beween PI and sperm motility or MI, respectively, in fertile group whereas there was no correlation in infertile group. These data suggest that SPA using the low temperature capacitation in TYB can be a valuable diagnostic tool for the assessment of male fertility in vitro and provide an important supplement to the traditional tests of sperm quality.