title: Resurrection of a bull by cloning from organs frozen without cryoprotectant in a -80 degrees c freezer for a decade.
abstract: Frozen animal tissues without cryoprotectant have been thought to be inappropriate for use as a nuclear donor for somatic cell nuclear transfer (SCNT). We report the cloning of a bull using cells retrieved from testicles that had been taken from a dead animal and frozen without cryoprotectant in a -80 degrees C freezer for 10 years. We obtained live cells from defrosted pieces of the spermatic cords of frozen testicles. The cells proliferated actively in culture and were apparently normal. We transferred 16 SCNT embryos from these cells into 16 synchronized recipient animals. We obtained five pregnancies and four cloned calves developed to term. Our results indicate that complete genome sets are maintained in mammalian organs even after long-term frozen-storage without cryoprotectant, and that live clones can be produced from the recovered cells.
authors.affiliation: Gifu Prefectural Livestock Research Institute, Takayama, Gifu, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Hoshino
authors.forename: Noboru
authors.initials: N
authors.lastname: Hayashi
authors.forename: Shunji
authors.initials: S
authors.lastname: Taniguchi
authors.forename: Naohiko
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Kenji
authors.initials: K
authors.lastname: Sakai
authors.forename: Tsuyoshi
authors.initials: T
authors.lastname: Otani
authors.forename: Akira
authors.initials: A
authors.lastname: Iritani
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Saeki
mesh_terms: D000818:Animals; D002417:Cattle; D019976:Cloning, Organism / Q000379:methods*; D015925:Cryopreservation; D003451:Cryoprotective Agents; D004624:Embryo Transfer / Q000379:methods; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D005615:Freezing; D008297:Male; D053652:Nuclear Transfer Techniques; D013737:Testis / Q000378:metabolism
chemical_list: D003451:Cryoprotective Agents