Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Light and Electron Microscopic Observation in the Frozen-thawed Mouse Testicular Tissues

동결보존된 생쥐 고환조직 세포의 광학 및 전자현미경적 관찰

  • Han, Sang-Chul (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine) ;
  • Song, Sang-Jin (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine) ;
  • Lee, Sun-Hee (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine) ;
  • Oh, Seung-Han (Department of Life Science, College of Natural Science Soonchunhyang University) ;
  • Koong, Mi-Kyung (Department of Obstetrics & Gynecology, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine) ;
  • Park, Yong-Seog (Laboratory of Reproductive Biology and Infertility, Samsung Cheil Hospital & Women's Healthcare Center, Sungkyunkwan University School of Medicine)
  • 한상철 (삼성제일병원 생식생물학 및 불임연구실) ;
  • 송상진 (삼성제일병원 생식생물학 및 불임연구실) ;
  • 이선희 (삼성제일병원 생식생물학 및 불임연구실) ;
  • 오승한 (순천향대학교 생명과학과) ;
  • 궁미경 (삼성제일병원 산부인과) ;
  • 박용석 (삼성제일병원 생식생물학 및 불임연구실)
  • Published : 2003.06.30
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Abstract

Objective: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. Methods: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. Results: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. Conclusions: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

Keywords

References

  1. Bourne H, Watkins W, Speirs A, Baker H. Pregnancies after intracytoplasmic injection of sperm collected by fine needle biopsy of the testis. Fertil Steril 1995; 64: 433-6 https://doi.org/10.1016/S0015-0282(16)57747-4
  2. Crister JK, Huse-Benda AR, Aaker DV, Arneson BW, Ball GD. Cryopreservation of human spermatozoa III. The effect of cryoprotectants on motility. Fertil Steril 1998; 50: 314-20
  3. Schoysman R, Vanderzwalmen P, Nijs M, Segal L, Segal-Bertin G, Geerts L, et al. Pregnancy after fretilization with human testicular spermatozoa. Lancet 1993; 342: 1237
  4. 박용석, 전진현, 변혜경, 김종현, 서주태, 이유식 등. 고환조직 정자채취술과 세포질내 정자주입술을 이용한 고환조직 정자의 수정률과 임신율. 대한불임학회지 1997; 24: 101-9
  5. Borini A, Sereni E, Bonu MA, Flamigni C. Freezing a few testicular spermatozoa retrieved by TESA. Mol Cell Endo 2000; 169: 27-32 https://doi.org/10.1016/S0303-7207(00)00347-6
  6. Oates RD, Mulhall J, Burgess C, Cummingham D, Carson R. Fertilization and pregnancy using intentionally cryopreserved testicular tissue as the sperm source for intracytoplasmic sperm injection in 10 men with non-obstructive azoospermia. Hum Reprod 1997; 12: 734-9 https://doi.org/10.1093/humrep/12.4.734
  7. Liu J, Nagi Z, Goossens A, Touraye H, Camus M, Van Steirteghem A, et al. Pregnancy after testicular sperm extraction and intracytoplasmic sperm injection non-obstructive azoospermia. Hum Reprod 1995; 10: 1457-60 https://doi.org/10.1093/HUMREP/10.6.1457
  8. Craft I, Sirigotis MT. Simplified recovery, preparation and cryopreservation of testicular spermatozoa. Hum Reprod 1995; 10: 1623-7 https://doi.org/10.1093/oxfordjournals.humrep.a136142
  9. 박용석, 전진현, 이호준, 강인수, 김종현, 이유식 등. 동결-융해 후 회수된 고환 정자와 세정관내정자의 수정능력과 효율성에 관한 연구. 대한불학회지 1998; 25: 171-7
  10. 박용석, 이형송, 변혜경, 염혜원, 송상진, 임천규 등. 폐쇄성 무정자증 환자의 신선고환조직 정자와 동결고환조직 정자의 운동성이 임신율에 미치는 영향. 대한불임학회지 2001; 28: 155-60
  11. Polge C, Smith AU, Parkes AS. Revival of spermatozoa after verification and dehydration at low temperature. Nature 1949; 164: 666-76 https://doi.org/10.1038/164666a0
  12. Freshney RI. Culture of animal cells: a manual of basic technique. Wiley-Liss, New York. 1994: 253-65
  13. Nogueira D, Bourgain G, Van VG, Steirteghem AC. Light and electron microscopic analhysis of human testicular spermatozoa and spermatids from frozen and thawed testicular biopsies. Hum Reprod 1999;14: 2041-9 https://doi.org/10.1093/humrep/14.8.2041
  14. Ben-Yosef D, Yogev L, Hauser R, Yavetz H, Azem F, Yovel I, et al. Testicular sperm retrieval and cryopreservation prior to initiating ovarian stimulation as the first line approach in patients with nonobstructive azoospermia. Hum Reprod 1999; 14:1794-801 https://doi.org/10.1093/humrep/14.7.1794