Figure 1.
One of Yasufuku's testicles frozen for 13 years.
The testicle was stored in a −80°C freezer for 10 years and then transferred to liquid nitrogen for 3 years. (A) Yasufuku's frozen testicle. (B) Part of the caput epididymis (arrow). (C) Spermatic cords that had been cut into three pieces. Scale bars represent 2 cm.
Figure 2.
Differential interference-contrast micrograph of cell populations established from Yasufuku's frozen testicles.
Cell lines A and B were used for SCNT after primary culture, and cell lines C and D were cryopreserved and then subcultured with five passages. (A) Cell line A, consisting of fibroblast-like cells. (B) Cell line B, consisting of epithelial-like cells. (C) Cell line C, consisting of fibroblast-like cells. (D) Cell line D, consisting of epithelial-like cells. Pregnancies were obtained from SCNT embryos that had been cloned from cells of line A (A) and line D (D). Scale bars represent 100 µm.
Figure 3.
Calves cloned from Yasufuku's frozen testicles.
(A) A male calf derived from Yasufuku's testicles was born on 30 November 2007. Parturition was induced by injection of prostaglandin F2α after 287 days of gestation and the recipient animal delivered this calf two days after induction. The calf's birth weight was 18.5 kg and he remains healthy at the time of writing. (B) A male calf, derived from a vitrified SCNT embryo, that was delivered by Caesarean section on 5 March 2008, after 286 days of gestation. The calf's birth weight was 47.5 kg; he died two days after birth. (C) Male calves derived from vitrified SCNT embryos. The calf with an ear tag “c95” was born on 22 July 2008, at 287 days of gestation. Its birth weight was 32 kg. The calf with an ear tag “c66” was born on 31 July 2008, at 288 days of gestation. Its birth weight was 30 kg. Parturitions were induced as described above. Both remain healthy at the time of writing.
Table 1.
Development of embryos cloned from cells of three different Yasufuku cell lines.
Table 2.
DNA microsatellite analysis in 13 polymorphic locia.
Table 3.
DNA microsatellite analysis in 18 polymorphic locia.