title: Assessment of freezing procedures for rat immature testicular tissue.
abstract: Fertility preservation has been included in the management of childhood cancer treatment. Cryopreservation of immature testicular tissue is the only available solution for pre-pubertal boys. Different freezing protocols have been developed in several species but without a clearly identified procedure. We tried to evaluate several protocols for cryopreservation of rat immature testicular tissue. Twelve different freezing protocols using different (i) cryoprotectant (dimethylsulphoxide [DMSO] or 1,2-propanediol [PROH]), (ii) cryoprotectant concentration (1.5M or 3M), (iii) equilibration time (30 or 60 min), (iv) equilibration temperature (4 °C or room temperature), (v) size of testicular fragment (7.5mg or 15 mg), (vi) package (straws or cryovials), were compared using cord morphological damage evaluation. A testicular tissue piece of 7.5mg cryopreserved in cryovial using 1.5M DMSO, an equilibration time of 30 min at 4 °C showed fewer morphological alterations than the other protocols tested. The selected freezing protocol was able to maintain rat immature testicular tissue architecture, functionality after testicular pieces organotypic culture, and could be proposed in a human application.
authors.affiliation: EA 4308 Spermatogenesis and Male Gamete Quality, Reproductive Biology Laboratory-CECOS, Rouen University Hospital, Institute for Biomedical Research, University of Rouen, Rouen, France. albanne.travers@aliceadsl.fr
authors.forename: A
authors.initials: A
authors.lastname: Travers
authors.forename: J P
authors.initials: JP
authors.lastname: Milazzo
authors.forename: A
authors.initials: A
authors.lastname: Perdrix
authors.forename: C
authors.initials: C
authors.lastname: Metton
authors.forename: A
authors.initials: A
authors.lastname: Bironneau
authors.forename: B
authors.initials: B
authors.lastname: Macé
authors.forename: N
authors.initials: N
authors.lastname: Rives
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide; D008297:Male; D051381:Rats; D013737:Testis / Q000033:anatomy & histology*; D046509:Tissue Culture Techniques / Q000662:veterinary; D014021:Tissue Preservation / Q000379:methods / Q000662:veterinary*
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide