Investigations of motility and fertilization potential in thawed cryopreserved mouse sperm from cold-stored epididymides

Toru Takeo, Kiyoko Fukumoto, Tomoko Kondo, Yukie Haruguchi, Yumi Takeshita, Yuko Nakamuta, Shuuji Tsuchiyama, Hidetaka Yoshimoto, Norihiko Shimizu, Ming Wen Li, Kristy Kinchen, Jadine Vallelunga, Kevin C K Lloyd, Naomi Nakagata

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Cold transport of epididymides from genetically modified mice is an efficient alternative to the shipment of live animals between research facilities. Mouse sperm from epididymides cold-stored for short periods can maintain viability. We previously reported that cold storage of mouse epididymides in Lifor® perfusion medium prolonged sperm motility and fertilization potential and that the sperm efficiently fertilized oocytes when reduced glutathione was added to the fertilization medium. Cryopreservation usually results in decreased sperm viability; an optimized protocol for cold storage of epididymides plus sperm cryopreservation has yet to be established. Here, we examined the motility and fertilization potential of cryopreserved, thawed (frozen-thawed) sperm from previously cold-stored mouse epididymides. We also examined the protective effect of sphingosine-1-phosphate (S1P) on sperm viability when S1P was added to the preservation medium during cold storage. We assessed viability of frozen-thawed sperm from mouse epididymides that had been cold-transported domestically or internationally and investigated whether embryos fertilized in vitro with these sperm developed normally when implanted in pseudo-pregnant mice. Our results indicate that frozen-thawed sperm from epididymides cold-stored for up to 48. h maintained high fertilization potential. Fertilization potential was reduced after cold storage for 72. h, but not if S1P was included in the cold storage medium. Live pups were born normally to recipients after in vitro fertilization using frozen-thawed sperm from cold-transported epididymides. In summary, we demonstrate an improved protocol for cold-storage of epididymides that can facilitate transport of genetically engineered-mice and preserve sperm viability after cryopreservation.

Original languageEnglish (US)
Pages (from-to)12-17
Number of pages6
Issue number1
StatePublished - Feb 2014


  • Cold storage
  • Cold transport
  • Epididymides
  • In vitro fertilization
  • Motility
  • Sperm
  • Sphingosine-1-phosphate

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)


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