title: Cryosurvival of isolated testicular cells and testicular tissue of tench Tinca tinca and goldfish Carassius auratus following slow-rate freezing.
abstract: Experiments were carried out to test the efficiency of cryopreservation of whole testicular tissue in tench Tinca tinca and goldfish Carassius auratus and compare it to cryopreservation of isolated testicular cells. Additionally, effects of three cryoprotectants (dimethyl sulphoxyde - Me2SO, methanol - MeOH and ethylene glycol - EG) at three concentrations (1M, 2M and 3M) on post-thaw cell viability were assessed. Tissue pieces/isolated testicular cells were diluted in cryomedia and cryopreserved by slow-rate freezing (1°C/min to -80°C followed by a plunge into the liquid nitrogen). In both species Me2SO and EG generally yielded higher cryosurvival of early-stage germ cells than MeOH, while spermatozoa of neither species displayed such a pattern. In most cases a 3M>2M>1M viability pattern emerged in both species for both sample types regardless of the cryoprotectant used. Sample type (dissociated testicular cells vs testicular tissue) did not seem to affect viability rates of tench early-stage germ cells and goldfish spermatozoa, while the opposite was observed for tench spermatozoa and goldfish early-stage germ cells. Additionally, through histological analysis we displayed that tissue structure mainly remained unaltered after thawing in goldfish. These results indicate that cryopreservation of whole testicular tissue is indeed a valid alternative method to cryopreservation of dissociated testicular cells. Early-stage germ cells obtained from cryopreserved testis can be further used in different purposes such as transplantation into suitable donors while viable sperm might be used for fertilization when feasible.
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary; University of Novi Sad, Department of Biology and Ecology, Trg Dositeja Obradovića 3, 21000 Novi Sad, Serbia. Electronic address: zoran.marinovic@dbe.uns.ac.rs.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Marinović
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary.
authors.forename: Jelena
authors.initials: J
authors.lastname: Lujić
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary.
authors.forename: Eszter
authors.initials: E
authors.lastname: Kása
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary.
authors.forename: Gergely
authors.initials: G
authors.lastname: Bernáth
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary.
authors.forename: Béla
authors.initials: B
authors.lastname: Urbányi
authors.affiliation: Szent István University, Department of Aquaculture, Páter K. u. 1., 2100 Gödöllő, Hungary.
authors.forename: Ákos
authors.initials: Á
authors.lastname: Horváth
mesh_terms: D000818:Animals; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D003530:Cyprinidae; D005306:Fertilization / Q000187:drug effects; D005615:Freezing; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000187:drug effects; D013737:Testis; D013997:Time Factors
chemical_list: D003451:Cryoprotective Agents
keywords: Germ cells; Spermatozoa cryopreservation; Testis cryopreservation