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1 정연경, 이정렬, 문정희, 김현준, 한상훈, 지병철, 구승엽, 서창석, 김석현, 최영민, 김정구, 문신용 大韓不妊學會誌 : 第 33 卷第 1 號 2006 Kor. J. Fertil. Steril., Vol. 33, No. 1, 2006, 3 무력정자증환자에서인간난포액과 SpermGrad 를이용한정자처리법의비교 서울대학교의과대학산부인과학교실 1, 분당서울대학교병원산부인과 2, 서울대학교의학연구원인구의학연구소 3 정연경 1 이정렬 1 문정희 2 김현준 2 한상훈 2 지병철 2 구승엽 1,3 서창석 1,2,3 김석현 1,3 최영민 1,3 김정구 1 문신용 1,3 Comparison between Human Follicular Fluid and SpermGrad for Sperm Preparation in Asthenozoospermia Youn Kyung Chung 1, Jung Ryeol Lee 1, Jeong-Hee Moon 2, Hyun-Jun Kim 2, Sang-Hoon Han 2, Byung-Chul Jee 2, Seung-Yup Ku 1,3, Chang-Suk Suh 1,2,3, Seok-Hyun Kim 1,3, Young-Min Choi 1,3, Jung-Gu Kim 1, Shin-Yong Moon 1,3 1Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Seoul, 2Department of Obstetrics and Gynecology, Seoul National University Bundang Hospital, Seongnam, 3Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Seoul, Korea Objective: We tested the usefulness of swim-down technique using human follicular fluid (hff) in sperm preparation. Methods: Semen samples were obtained from twelve male partners showing asthenozoospermia (sperm motility < 50%) at the time of routine andrologic evaluation in Seoul National University Bundang Hospital. After dividing into two aliquots, each samples were processed either by swim-down using 100% hff or density gradient using SpermGrad. Sperm quality was assessed by computer-assisted semen analyzer (CASA). Results: Motility, Rapid motility, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement), and hyperactivated sperms were significantly increased, and LIN (mean linearity) was decreased significantly after sperm preparation in both groups. Motility was significantly higher after swim-down using 100% hff when compared with density gradient using SpermGrad (81.2±4.7 vs. 67.6±2.3, p=0.02). The other parameters assessed by CASA were not different between the two methods. Conclusion: Swim-down method with 100% hff may be a useful method in preparation of sperm from asthenozoospermia. Key Words: Asthenozoospermia, Density gradient, Human follicular fluid, SpermGrad, Swim-down 체외수정시술 (in vitro fertilization, IVF) 이나인공수정 (intrauterine insemination, IUI) 등의보조생식술 (assisted reproductive technology, ART) 을실시하기위해서는정액에서정장 (seminal plasma) 을신속히제거하고운동성이있는정자를효과적으로분리하 는과정이필수적이다. 이상적인정자처리법은정상형태의운동성정자를최대한많이회수할수있어야하고, 첨체반응 (acrosome reaction) 을높여수정률을증가시킬수있어야하며, 정장, 세균, 백혈구등을완전히제거할수있어야한다. 1 주관책임자 : 서창석, 우 ) 경기도성남시분당구구미동 300 번지, 분당서울대학교병원산부인과 Tel: (031) , Fax: (031) , suhcs@snu.ac.kr

2 무력정자증환자에서인간난포액과 SpermGrad 를이용한정자처리법의비교 정자처리법은현재까지 10여가지이상이개발되어있으며이중가장보편적으로사용되는방법은 swim-up 방법과밀도차분리법 (density gradient separation technique) 이다. Swim-up 방법은비교적정액처리과정이간단하고정액양, 정자수, 정자의운동성이정상인경우에는효율적이어서많이사용되고있으나, 정자감소증 (oligozoospermia), 정자무력증 (asthenozoospermia) 과같은비정상정액인경우에는그효용성이낮은것으로보고되고있다. 2 한편, 밀도차분리법은비정상정액상을보이는정액군에서도정자처리효과가높은것으로알려져있으며, 3 현재여러불임센터에서남성인자가있는불임환자의정액을처리할때는밀도차분리법을주로사용하고있다. 밀도차분리법중에서 Percoll (Pharmacia, Sweden) 용액을이용한결과와효용성에대한보고도많았으나현재는사용되지않고있다. Percoll 은 polyvinylpyrollidone (PVP) 으로코팅된 silica particle로서 PVP 자체에다소독성이있고 NaCl이첨가된배양액내에서 silica로부터결합이느슨해지는경향이있어결국에는 silica gel이정자두부의 DNA나운동성에악영향을줄수있는것으로알려져있다. 이후 Percoll의장점을가지면서독성이약한대체재로개발된 SpermGrad (VitroLife, Sweden) 와 Puresperm (Nidacon, South Africa) 등이현재많이사용되고있다. 정자의운동성은특히인공수정에서성공적인임신에가장중요한요소중하나이므로정자무력증을가진환자의정액을처리하는방법에대한연구가이전부터다양하게이루어져왔다. 인간의난포액 (human follicular fluid, hff) 을정자처리에사용하여정자의수정능력을높이려는연구는대부분 hff를배양액에첨가하여그결과를살펴본것이며, 직접적인정자분리법에적용한것은 hff를이용한 swim-up법과밀도차분리법을비교한것이고, swimdown법을이용한정자분리법에대한연구는미비한실정이다. 4~6 이에저자들은정자무력증을보이는정액을대상으로 hff를이용한 swim-down법과 SpermGrad를이용한밀도차분리법을적용하였을때정자의운동성에미치는영향을비교분석함으로써정자처리법에서 hff의효용성에대한검토와각검사법간의 상호보완성여부를알아보기위하여본연구를시행하였다. 연구대상및방법 1. 연구대상 2004년 3월부터 7월까지분당서울대학병원산부인과를방문하여기본적불임검사로서정액검사를시행한환자중정자의운동성이 50% 미만으로세계보건기구 (World Health Organization, WHO) 의정의상정자무력증의소견을보인 12명을연구대상으로하였다. 7 12명의남성의평균연령은 33.3±2.9세이었으며처리전정자의평균운동성은 41.8±4.6% ( 범위 : 30~46%), 평균농도는 6.4± /ml이었다. 2. 연구방법 1) 정액검사와정자처리 48시간의금욕후에수음을통해얻어진정액을최소 30분간액화시킨후기본적인정액성상을확인하기위해 10 µl를취하여컴퓨터정액분석기 (computer-assisted semen analyzer, SAIS, Semen Analysis Image System version 10.1, Medical Supply, Korea) 를이용한정액분석을시행하여농도, 운동성, 빠른운동성정자의비율, straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), beat/ cross frequency (BCF), straightness [STR, (VSL/VAP) 100], mean linearity [LIN, (VSL/VCL) 100] 및과활동성정자 (hyperactivated sperm) 의비율에관한자료를얻었다. 컴퓨터정액분석기의설정은다음과같았다 : Image acquisition rate 30 frame/sec, tract sampling time 1 sec, minimum sampling for motility 5 frames, minimum sampling for velocity 10 frames, minimum sampling for ALH 12 frames, maximum velocity 250 µm/s, minimum velocity 10 µm/s, cell size range 10~250 pixels. 과활동성정자의기준은 VCL > 80 µm/s, LIN < 65%, ALH > 6.5 µm/s으로정하였다. 8 이후정액은 modified Ham's F-10 (Gibco, Grand Island, USA) 배양액으로 2회세척하였다. 즉, 3 ml 의 modified Ham's F-10으로희석한후 3,000 rpm으로 5분간원심분리하여상층액을제거하고정자괴

3 정연경, 이정렬, 문정희, 김현준, 한상훈, 지병철, 구승엽, 서창석, 김석현, 최영민, 김정구, 문신용 Raw sperm + Ham's * Debris + dead sperm Others & dead sperm or Isolated sperm SpermGrad hff Live sperm with good quaility Pellet collection Remove sup. Figure 1. Procedures of sperm preparation. *: Rinse and resuspension, : Centrifuge for 30 seconds (* 2), : Centrifuge for 15 minutes or incubation for 1 hours (sperm pellet) 를다시 3 ml의 modified Ham's F-10으로희석한후 3,000 rpm으로 30초 spin down하였다. 이후운동성정자가풍부한상층액을 2 등분하여 100% hff 1.0 ml이담긴튜브와 100% 등장성 SpermGrad (VitroLife, Sweden) 1.0 ml이담긴튜브에각각조심스럽게얹었다 (Figure 1). 100% hff를이용한 swim-down법에서는 5% CO 2 배양기안에서 37 의온도로 60분간배양한후상층액은제거하고정자괴만을얻어 10% Synthetic Serum Substitute (SSS, Irvine Scientific, Santa Ana, USA) 를포함한 modified Ham's F-10으로 2회세척한후상층액을제거하고정자괴를취하여컴퓨터정액분석을시행하였다. SpermGrad 를이용한밀도차분리법에서는 3,000 rpm으로 15분간원심분리한후상층액은제거하고남은정자괴를 10% SSS를포함한 modified Ham's F-10으로 2회세척후상층액은제거하고남은정자괴를취하여컴퓨터정액분석을시행하였다

4 무력정자증환자에서인간난포액과 SpermGrad 를이용한정자처리법의비교 Table 1. Parameters of human sperms after preparation with swim-down using 100% hff and density gradient using SpermGrad Fresh ejaculate Swim-down using 100% hff Density gradient using SpermGrad Total concentration ( 10 8 ) 19.8± ±1.2 a 7.8±1.8 Motility (%) 41.8± ±4.7 a, 67.6±2.3 a Motility, rapid (%) 28.4± ±6.6 a 47.4±7.0 a VCL (µm/s) 82.5± ±7.7 a 108.4±7.7 a VSL (µm/s) 29.5± ± ±2.0 VAP (µm/s) 41.0± ± ±3.5 ALH (µm) 3.7± ±0.4 a 4.9±0.3 a BCF (Hz) 16.1± ± ±0.8 STR (%) 72.4± ± ±1.4 LIN (%) 37.1± ±2.0 a 34.8±2.0 a Hyperactivated sperm (%) 1.4± ±5.1 a 5.3±5.5 a n = 12. Data present: mean ± SEM except, : mean ± SD. a: p<0.05 when compared with fresh ejaculate (Mann-Whitney U test). : p<0.05 when compared with density gradient using SpermGrad (Mann-Whitney U test). hff: human follicular fluid, VCL: curvilinear velocity, VSL: straight-line velocity, VAP: average path velocity, ALH: amplitude of lateral head displacement, BCF: beat/cross frequency, STR: straightness, LIN: mean linearity 2) 난포액의수집분당서울대학교병원산부인과에서체외수정시술을시행하는여성에서난자채취시얻은흡인액중난자를선별하고이때난포액을수집하였다. 수집한 hff는세포파편등을제거하기위해 5.0 µm, 0.8 µm, 0.2 µm pore size의필터로연속적으로여과처리한후 56 water bath에서 30분간처리하여비활성화 (inactivation) 시켰다. 이후정자처리사용전까지 -70 에서보관하였으며사용전 37 에서해동한후 0.22 µm filter에여과하여사용하였다. 3) 통계분석연구결과에대한통계학적인분석은 SPSS for Windows (version 12.0, SPSS Inc., Chicago, USA) 프로그램을이용하였다. 두가지정자처리방법에따른변수들의차이에대해서는 Mann-Whitney U test 를사용하였으며양측검정에서 p<0.05인경우통계학적인유의성이있는것으로판정하였다. 결과처리전정액과비교할때 100% hff를이용한 swim-down법과 SpermGrad를이용한밀도차분리법모두에서운동성과빠른운동성정자의비율, VCL, ALH 치및과활동성정자의비율이통계적으로유의하게상승하였고, LIN 치는유의하게감소하였다. 그러나 VSL, VAP, BCF, STR 치는처리후에도변화가없었다 (Table 1). 100% hff를이용한 swim-down법과 SpermGrad 를이용한밀도차분리법을상호비교시운동성만이 100% hff를이용한 swim-down에서 SpermGrad 를이용한밀도차분리법에비하여통계적으로유의하게높게나타났다 (81.2±16.3 vs. 67.6±7.9 p=0.02). 빠른운동성정자의비율, VCL, VSL, VAP, ALH, BCF, STR, LIN 치및과활동성정자의비율에는모두유의한차이가없는것으로나타났다. 고찰본연구결과 100% hff를사용하여 swim-down 법으로정자를처리하였을때 SpermGrad를이용한밀도차분리법과마찬가지로정자처리를시행하지않은상태의정액에서보다운동성과빠른운동성

5 정연경, 이정렬, 문정희, 김현준, 한상훈, 지병철, 구승엽, 서창석, 김석현, 최영민, 김정구, 문신용 정자의비율, VCL, ALH 치및과활동성정자의비율이유의하게상승하였으며, SpermGrad 를이용한밀도차분리법에비해서운동성이유의하게증가하여양질의정자를회수하는데 SpermGrad를이용한밀도차분리법를대체할수있는우수한방법으로생각된다. 컴퓨터정액분석의지표와임신결과와의관계에대한기존의연구결과들을살펴보면, Hirano 등 9 은농도, 운동성, 모양, ALH, VCL, VSL 치, 빠른운동성정자의비율등이정자의수정능력과관계가있다고하였고, Donnelly 등 10 은운동성, VAP, VSL, VCL, LIN 치와과활동성정자의비율이수정율과관계가있다고하였다. 또한 Geyter 등 11 은 VCL, ALH, VAP, VSL 치가수정률과유의한관계가있으며그중에서도 VCL이가장뚜렷한관계가있다고하였고, 과활동성정자의비율또한수정과유의한관계가있다고하였다. Fabbri 등 12 은 hff를포함한배양액에서정자를배양하였을때정자의운동성, VCL, ALH 치가증가하였다고하였으며, Bajamonte 등 4 도심한정자이상을동반하는환자의정액을 Percoll과 50% hff를혼합하여 swim-up 방법으로처리하였을때수정능과환자당배아이식비율및임신율의증가를보고하여체외수정시술시심한정자이상이있는경우 hff의이용이매우유용하다고보고하였다. 보조생식술에서정액처리나수정시배양액에 hff를포함하는것이임신결과에어떤영향을주는지에대해서는아직논란이있는실정이다. hff 를포함한배양액을사용하여체외수정시술시그결과가향상되었다는보고도있으며, 4,21 Hong 등 22 도 hff가정자의운동성을증가시키고첨체반응을증대시키며난자쪽으로이동하여투명대에부착하여이를용해하여난자내로진입하는과정을촉진시킴으로보조생식술의결과를향상시킨다고하였다. 그러나배양액에 hff를포함하였을때정자가난자와반응하는능력을감소시켰다고다른보고들도있으므로 23~25 이에대해서는좀더연구가필요하리라생각된다. 한편, 본연구의결과를보조생식술시행시정자처리에곧바로적용하기에는몇가지제한점이있다. 첫째, 연구에포함된대상군의수가 12명으로전체 군을반영하기에는어려움이있다. 따라서향후대상군의집단을더욱크게하여본연구의결과에대한확증이필요할것으로사료된다. 둘째본연구결과 100% hff를사용하여 swim-down법으로정자를처리하였을때 SpermGrad를이용한밀도차분리법과마찬가지로운동성을증가시킬수있었으며 SpermGrad를이용한밀도차분리법에비해서도운동성이유의하게증가하였으나 SpermGrad 군이 hff 군보다총정자의수가많으므로운동성을가지는정자의총수는두군간에유의한차이는보이지않았다. 수음을통해얻어진정액을전처치후같은양으로나누어 SpermGrad와 hff로정자처리를하였을때각각의정자처리후두군간에총정자수의차이를보이는것에대해서 ( 통계적으로유의하지는않았다.) hff 군에서배양시간에따른영향여부등에대한비교분석이추가로필요할것으로사료된다 (Table 1). 셋째, 본연구는두군간에보조생식술에서정자처리에대한결과를비교한것으로그비교분석대상이정액검사에제한되어있다. 두군에서다른방법으로처리한정액을이용하여시행한보조생식술의결과, 즉수정률, 배아의발달, 임신율등의변화에대한자료가없어실제임상에서의유용성등에대한자료가없다. 향후대상군집단을크게하고이를보조생식술에이용하여그결과, 즉수정율, 임신율등에주는영향에대한연구가필요하겠다. 본연구에서는 hff를배양액으로이용하여 swimdown법을적용하였다. 현재개발된정액처리법중가장보편적으로사용되고있는방법은 swim-up 방법과밀도차분리법인데이중 swim-up법은비교적정액처리과정이간단하고운동성이있으며정상적인형태를가진정자의회수에효율적이어서많이사용되고있으나, 희소정자증, 무력정자증과같은남성불임에서얻어진정액인경우에는그효용성이낮은것으로보고되고있다. 2 그러므로남성불임이있는환자의정액을처리할경우에는정자의회수율을높이기위해밀도차분리법이많이이용되고있다. 본연구에서는정자감소증에서많이사용되고있는밀도차분리법과 hff를이용한 swim-down 방법을비교하였는데대부분의논문에서는밀도차분리법과 swim-up 방법을서로비교분석하였다. 이에 hff를

6 무력정자증환자에서인간난포액과 SpermGrad 를이용한정자처리법의비교 배양액으로이용하는정자처리법의선택에있어서 swim-down과다른방법, 예를들어 swim-up, swimcross법등을비교하여각각의방법에대한분석이필요하겠다. 본연구결과정자무력증을보이는정액에서는 100% hff를사용하여 swim-down법으로정자를처리하였을때 SpermGrad를이용한밀도차분리법과마찬가지로운동성을증가시킬수있었으며 Sperm- Grad를이용한밀도차분리법에비해서도운동성이더욱유의하게증가하였다. 따라서 100% hff를사용한 swim-down법을양질의정자를회수하는데 SpermGrad를이용한밀도차분리법의대체방법으로제시할수있다고사료된다. 경제성과배양액에의한정자의손상, 독성여부등을고려할때인간난포액이 SpermGrad 등의대체물질로사용될수있을것으로생각된다. 그러나인간난포액의감염여부와다른개체에서의추출등에따른항원-항체반응등에대한고려가필요하며, 향후실제임상에서의유용성에대한추가연구가수행되어야한다. 참고문헌 1. Berger RE, Karp LE, Williamson RA, Koehler J, Moor DE, Holmes LK. The relationship of pyrospermia and seminal fluid bacteriology to sperm function as reflected in the sperm penetration assay. Fertil Steril 1982; 37: Ohashi K, Saji F, Wakimoto A, Kato M, Tsutsui T, Tanizawa O. Preparation of oligozoospermic and/or asthenozoospermic semen for intrauterine insemination using the SpermPrep semen filtration column. Fertil Steril 1992; 57: Jaroudi KA, Carver-Ward JA, Hamilton CJ, Sieck UV, Sheth KV. Percoll semen preparation enhances human oocyte fertilization in male-factor infertility as shown by a randomized cross-over study. Hum Reprod 1993; 8: Bajamonte M, Ruvolo G, Cimino C, Cittadini E. Advantages of treatment with human follicular fluid in the management of severely dyspermic patients in human in vitro fertilization programs. Acta Eur Fertil 1994; 25: Ruvolo G, Cassara G, Di Bella MA, Parrinello D, Manno M, Cimino C, et al. Activation of the acrosome reaction after treatment with human follicular fluid. A morphofunctional evaluation useful for in vitro fertilization. Acta Eur Fertil 1995; 26: Chao HT, Ng HT, Kao SH, Wei YH, Hong CY. Human follicular fluid stimulates the motility of washed human sperm. Arch Androl 1991; 26: World Health Organization. WHO Laboratory Manual for the Examination of Human Semen and Sperm- Cervical Mucus Interaction, 4th ed. Cambridge: Cambridge University Press, Murad C, De Lamirande E, Gagnon C. Hyperactivated motility is coupled with interdependent modifications at axonemal and cytosolic levels in human spermatozoa. J Androl 1992; 13: Hirano Y, Shibahara H, Obara H, Suzuki T, Takamizawa S, Yamaguchi C, et al. Relationships between sperm motility characteristics assessed by the computer-aided sperm analysis (CASA) and fertilization rates in vitro. J Assist Reprod Genet 2001; 18: Donnelly ET, Lewis SE, McNally JA, Thompson W. In vitro fertilization and pregnancy rates: the influence of sperm motility and morphology on IVF outcome. Fertil Steril 1998; 70: De Geyter C, De Geyter M, Koppers B, Nieschlag E. Diagnostic accuracy of computer-assisted sperm motion analysis. Hum Reprod 1998; 13: Fabbri R, Porcu E, Lenzi A, Gandini L, Marsella T, Flamigni C. Follicular fluid and human granulosa cell cultures: influence on sperm kinetic parameters, hyperactivation, and acrosome reaction. Fertil Steril 1998; 69: Bongso A, Ng SC, Fong CY, Ratnam S. Cocultures: a new lead in embryo quality improvement for assisted reproduction. Fertil Steril 1991; 56: de Lamirande E, Leclerc P, Gagnon C. Capacitation as a regulatory event that primes spermatozoa for the acrosome reaction and fertilization. Mol Hum Roprod

7 정연경, 이정렬, 문정희, 김현준, 한상훈, 지병철, 구승엽, 서창석, 김석현, 최영민, 김정구, 문신용 1997; 3: Coddington CC, Franken DR, Burkman LJ, Oosthuizen WT, Kruger T, Hodgen GD. Functional aspects of human sperm binding to the zona pellucida using the hemizona assay. J Androl 1991; 12: Yao Y, Ho P, Yeung WS. Effects of human follicular fluid on the capacitation and motility of human spermatozoa. Fertil Steril 2000; 73: Hamamah S, Lanson M, Barthelemy C, Garrigue MA, Muh JP, Royere D, et al. Analysis of the lipid content and the motility of human sperm after follicular fluid treatment. Andrologia 1995; 27: Meizel S, Pillai MC, Diaz-Perez E, Thomas P. Initiation of the human sperm acrosome reaction by components of human follicular fluid and cumulus secretions including steroids. In: Bavister BD, Cummins J, Roldan ERS, eds. Fertilization in mammals. Norwell, MA, USA: Serono Symposia, 1990: Morales P, Llanos M, Gutierrez G, Kohen P, Vigil P, Vantman D. The acrosome reaction-inducing activity of individual human follicular fluid samples is highly variable and is related to the steroid content. Hum Reprod 1992; 7: Burrello N, Vicari E, D'Amico L, Satta A, D'Agata R, Calogero AE. Human follicular fluid stimulates the sperm acrosome reaction by interacting with the gamma-aminobutyric acid receptors. Fertil Steril 2004; 82: Giorgetti C, Hans E, Spach JL, Auquier P, Roulier R. In-vitro fertilization in cases with severe sperm defect: use of a swim-across technique and medium supplemented with follicular fluid. Hum Reprod 1992; 7: Hong CY, Chao HT, Lee SL, Wei YH. Modification of human sperm function by human follicular fluid: a review. Int J Androl 1993; 16: Yao YQ, Chiu CN, Ip SM, Ho PC, Yeung WS. Glycoproteins present in human follicular fluid that inhibit the zona-binding capacity of spermatozoa. Hum Reprod 1998; 13: Qiao J, Yeung WS, Yao YQ, Ho PC. The effects of follicular fluid from patients with different indications for IVF treatment on the binding of human spermatozoa to the zona pellucida. Hum Reprod 1998; 13: Munuce MJ, Caille AM, Botti G, Berta CL. Modulation of human sperm function by follicular fluid. Andrologia 2004; 36:

8 무력정자증환자에서인간난포액과 SpermGrad 를이용한정자처리법의비교 = 국문초록 = 목적 : 보조생식술에서의정자처리법으로서인간난포액을이용한 swim-down 방법의유용성을확인해보고자하였다. 연구방법 : 분당서울대학교병원산부인과에서불임평가목적으로정액검사를시행할때정자무력증 (asthenozoospermia, sperm motility < 50%) 을보이는 12명의남성을대상으로하였다. 이들에서검사후남은정액을 100% 인간난포액을이용한 swim-down법과 SpermGrad를이용한밀도차분리법을적용하여각각처리하고컴퓨터정액분석기를이용하여정액검사를시행하였다. 결과 : 두군모두에서운동성과빠른운동성정자의비율, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement) 치및과활동성정자의비율이통계적으로유의하게상승하였고 LIN (mean linearity) 치는유의하게감소하였다. 100% 인간난포액을이용한 swim-down법에서정자의운동성이 SpermGrad를이용한밀도차분리법에비하여유의하게높게나타났으며 (81.2±4.7 vs. 67.6±2.3, p=0.02), 다른변수들은두군에서차이를보이지않았다. 결론 : 100% 인간난포액을이용한 swim-down법은정자무력증을보이는경우유용한정자처리법으로사료된다. 중심단어 : 정자무력증, 밀도차분리법, 인간난포액, SpermGrad, Swim-down

Microsoft Word - 3-서창석.DOC

Microsoft Word - 3-서창석.DOC 대한생식의학회지 : 제 34 권제 3 호 2007 분당서울대학교병원산부인과 1, 서울대학교의과대학산부인과학교실 2 김용진 1 지병철 1 서창석 1,2 김석현 2 Effects of Conversion of Infertility Treatment on Semen Quality Yong Jin Kim 1, Byung Chul Jee 1, Chang Suk Suh