Abstract
In place of gametes, ovaries can be used as an alternative resource for cryopreservation. At the previous IETS meeting, we showed that in the Syrian hamster, pups could be successfully produced through the transplantation of fresh ovaries. In addition, the immunotolerance of interstrain ovarian transplantation was described. In this study, we attempted to produce pups via transplantation of cryopreserved ovaries to foster mothers. Our goal was to establish a preservation method for hamster strains based on ovarian cryopreservation in combination with ovarian transfer. Ovaries were cryopreserved by vitrification according to methods described for mouse ovaries (Migishima F et al. 2003 Biol. Reprod. 68, 881–887). In brief, ovaries were collected from 2- to 3-week-old HAW strain (white coat) females. Each ovary was cut into 4 sections, placed into 1 tube, and cryopreserved by vitrification using DAP213 solution (2 m dimethyl sulfoxide, 1 m acetamide, and 3 m propylene glycol; Nakagata N 1989 J. Reprod. Fertil. 87, 479–483). Thawed ovaries were transferred into the ovarian bursae (2 ovarian sections per ovarian bursa) of 3-week-old recipient females with agouti coat (Slc:Syrian) under anesthesia with isoflurane. Half of the recipients’ ovaries were removed immediately before transfer; i.e. both donor and recipient ovaries were present in each ovarian bursa. Successful ovarian transplantation was verified by examining the coat color (non-agouti color) of offspring produced in test matings between mature recipients and Slc:Syrian males. Out of 6 recipients tested, 2 became pregnant and delivered pups. According to the coat color of the pups, 1 of the 2 recipients delivered a pup produced from transplanted ovaries (1 out of 4 pups). Regardless of this low success rate, our results indicate that pups can be produced from cryopreserved ovaries via interstrain transplantation of ovaries to foster mothers in the Syrian hamster. Thus, ovarian cryopreservation by vitrification is a promising strategy for strain preservation of the Syrian hamster. This work was supported by a grant from the Ministry of Health, Labor, and Welfare, Japan.
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