Establishment of a strain of haemophilia-A pigs by xenografting of foetal testicular tissue from neonatally moribund cloned pigs

Sci Rep. 2017 Dec 5;7(1):17026. doi: 10.1038/s41598-017-17017-6.

Abstract

Grafting of testicular tissue into immunodeficient mice makes it possible to obtain functional sperm from immature donor animals that cannot be used for reproduction. We have developed a porcine model of human haemophilia A (haemophilia-A pigs) by nuclear transfer cloning from foetal fibroblasts after disruption of the X-linked coagulation factor VIII (F8) gene. Despite having a recessive condition, female F8+/- cloned pigs died of severe bleeding at an early age, as was the case for male F8-/Y cloned pigs, thus making it impossible to obtain progeny. In this study, therefore, we produced sperm from F8-/Y cloned pigs by grafting their foetal testicular tissue into nude mice. Two F8+/- female pigs were generated from oocytes injected with xenogeneic sperm. Unlike the F8+/- cloned pigs, they remained asymptomatic, and delivered five F8-/Y and four F8+/- pigs after being crossed with wild-type boars. The descendant F8-/Y pigs conserved the haemophilia phenotype. Thus, the present F8+/- pigs show resolution of the phenotypic abnormality, and will facilitate production of F8-/Y pigs as founders of a strain of haemophilia-A pigs for the development of new therapeutics for haemophilia A. This strategy will be applicable to other genetically modified pigs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Cloning, Organism / methods*
  • Factor VIII / genetics*
  • Female
  • Fetus*
  • Hemophilia A / genetics
  • Hemophilia A / metabolism
  • Hemophilia A / pathology*
  • Male
  • Mice
  • Mice, Nude
  • Nuclear Transfer Techniques*
  • Pregnancy
  • Swine
  • Testis / transplantation*
  • Transplantation, Heterologous

Substances

  • Factor VIII