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대한생식의학회지 제27권 제3호 2010년

고환조직 동결-융해 후 회수된 고환 정자에 대한 Hypo-osmotic Swelling (HOS) Test의 효과

성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실;성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실;성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실;성균관대학교 의과대학 삼성제일병원 생식생물학 및 불임연구실;성균관대학교 의과대학 삼성제일병원 산부인과;성균관대학교 의과대학 삼성제일병원 비뇨기과;

박용석;이형송;송상진;김정욱;강인수;서주태;,

Effect of Hypo-osmotic Swelling (HOS) Test on Subsequent Post-thaw Testicular Spermatozoa

Park, Yong-Seog;Lee, Hyoung-Song;Song, Sang-Jin;Kim, Jeong-Wook;Kang, Inn-Soo;Seo, Ju-Tae;

Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;Laboratory of Reproductive Biology & Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;Department of OB/GYN, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;Department of Urology, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine;

Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

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