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대한생식의학회지 제27권 제3호 2010년
고환조직 동결-융해 후 회수된 고환 정자에 대한 Hypo-osmotic Swelling (HOS) Test의 효과
성균관대학교 외과대학 삼성제일병원 생식생물학 및 불임연구실, 산부인과1, 비뇨기과2
박용석, 이형송, 송상진, 김정욱, 강인수1, 서주태2,
Effect of Hypo-osmotic Swelling (HOS) Test on Subsequent Post-thaw Testicular Spermatozoa
Yong-Seog Park, Hyoung-Song Lee, Sang Jin Song, Jeong-Wook Kim, Inn Soo Kang1, Ju Tae Seo2
Laboratory of Reproductive Biology & Infertility, Department of OB/GYN2, Department of Urology2, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine3, Seoul, Korea
Objectives: We have pervious reported that thawed testicular sperm and 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 LN2 and letting them at room temperature for 10 min followed by 37℃ 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 azoospermic 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.
키워드 : TESE, Seminiferous tubule, HOS test, Eosin-Y
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