An optimized method for cryogenic storage of Xenopus sperm to maximise the effectiveness of research using genetically altered frogs

Esther Pearl, Sean Morrow, Anna Noble, Adelaide Lerebours, Marko Horb, Matthew Guille

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Cryogenic storage of sperm from genetically altered Xenopus improves cost effectiveness and animal welfare associated with their use in research; currently it is routine for X. tropicalis but not reliable for X. laevis. Here we compare directly the three published protocols for Xenopus sperm freeze-thaw and determine whether sperm storage temperature, method of testes maceration and delays in the freezing protocols affect successful fertilisation and embryo development in X. laevis. We conclude that the protocol is robust and that the variability observed in fertilisation rates is due to differences between individuals. We show that the embryos made from the frozen-thawed sperm are normal and that the adults they develop into are reproductively indistinguishable from others in the colony. This opens the way for using cryopreserved sperm to distribute dominant genetically altered (GA) lines, potentially saving travel-induced stress to the male frogs, reducing their numbers used and making Xenopus experiments more cost effective.
Original languageEnglish
Pages (from-to)149-155
Number of pages7
Early online date17 Jan 2017
Publication statusPublished - 1 Apr 2017


  • RCUK
  • NERC
  • NE/018867/1
  • BB/K019988/1
  • Xenopus
  • sperm
  • cryopreservation
  • stock centres
  • genetically altered lines
  • 3Rs


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