title: Effects of various cryopreservation media and freezing-thawing on the morphology of rat testicular biopsies.
abstract: Currently, testicular sperm extraction is successfully combined with intracytoplasmic sperm injection into the oocyte (ICSI). Several pieces of a testicular biopsy can be frozen and thawed until the ICSI attempt. In this study, the effects of freezing-thawing on the morphology of rat testicular biopsies stored in different cryopreservation media were analysed. Each cryopreservation medium contained glycerol and/or dimethyl sulfoxide (DMSO) as cryoprotectants. In general, both glycerol and DMSO, when applied at moderate concentrations (6-25%), preserved the structure of the seminiferous epithelium. The freezing-thawing procedure had no significant effect on tubular diameter; however, it caused a 'folding' of the lamina propria and notable damage to Sertoli cells, spermatogonia and spermatocytes. Round and elongated spermatids and spermatozoa displayed occasional nuclear damage, vacuolization, and shrinkage/swelling of the cytoplasm. However, the vast majority of these cells maintained their normal structure in nearly all the applied cryomedia. It is concluded that freezing-thawing of testicular biopsies, and the cryopreservation medium, have a significant impact on the structure of the seminiferous epithelium, particularly on its basal compartment.
authors.affiliation: Department of Histology and Embryology, Medical School University of Zagreb, Croatia. davor@mef.hr
authors.forename: D
authors.initials: D
authors.lastname: Jezek
authors.forename: W
authors.initials: W
authors.lastname: Schulze
authors.forename: S
authors.initials: S
authors.lastname: Kalanj-Bognar
authors.forename: Z
authors.initials: Z
authors.lastname: Vukelić
authors.forename: V
authors.initials: V
authors.lastname: Milavec-Puretić
authors.forename: I
authors.initials: I
authors.lastname: Krhen
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005615:Freezing; D005990:Glycerol / Q000494:pharmacology; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D012662:Semen Preservation / Q000379:methods*; D012671:Seminiferous Tubules / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D012708:Sertoli Cells / Q000166:cytology / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects / Q000473:pathology / Q000648:ultrastructure; D013737:Testis / Q000187:drug effects* / Q000473:pathology / Q000648:ultrastructure
chemical_list: D003451:Cryoprotective Agents; D005990:Glycerol; D004121:Dimethyl Sulfoxide