Porcine embryos produced after intracytoplasmic sperm injection using xenogeneic pig sperm from neonatal testis tissue grafted in mice
Ali Honaramooz A C , Xiang-Shun Cui B , Nam-Hyung Kim B and Ina Dobrinski A DA Center for Animal Transgenesis and Germ Cell Research, Department of Clinical Sciences, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA 19348, USA.
B Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, South Korea.
C Present address: Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK S7N 5B4, Canada.
D Corresponding author. Email: dobrinsk@vet.upenn.edu
Reproduction, Fertility and Development 20(7) 802-807 https://doi.org/10.1071/RD08093
Submitted: 4 May 2008 Accepted: 3 July 2008 Published: 1 August 2008
Abstract
Embryo development after homologous intracytoplasmic sperm injection (ICSI) with sperm from testis tissue xenografts from pigs or any other farm animal species has not been evaluated critically. Here, we report development of porcine embryos in vitro following ICSI with sperm retrieved from xenografted neonatal pig testis. Small pieces of testis tissue from newborn piglets were grafted under the back skin of castrated immunodeficient mice (n = 4) and the xenografts were collected 8 months after grafting. Spermatozoa were recovered by mincing of the grafted tissue. For comparison, testicular, epididymal and ejaculated spermatozoa were also collected from mature boars. Oocytes injected with xenogeneic spermatozoa were either fixed to determine fertilisation processes (n = 89 in five replicates) or allowed to develop in vitro (n = 143 in four replicates). Xenogeneic porcine spermatozoa were fertilisation competent (24% v. 58%, 68%, 62% or 0% for xenogeneic v. control testicular, epididymal and ejaculated spermatozoa or no spermatozoa, respectively) and embryos developed to the blastocyst stage (8% v. 22%, 27%, 25% or 0%, respectively). These results demonstrate that porcine spermatozoa derived from immature testis tissue xenografted into mice are fertilisation competent, albeit at a lower rate than testicular, epididymal or ejaculated spermatozoa from control boars, and support embryo development after ICSI.
Acknowledgements
The authors thank Janet Turpin and Terry Jordan for animal care. This study was supported by National Research Initiative Competitive Grant no. 2003–35203–13486 (to I.D.) from the USDA Cooperative State Research, Education, and Extension Service and by grant no. 2R01 RR17359–06 (to I.D.) from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH). The contents of this paper are solely the responsibility of the authors and do not necessarily represent the official views of the NCRR or NIH. This study was also supported by a grant from the BioGreen 21 program, RDA, Republic of Korea (to N.H.K.).
Arregui, L. , Rathi, R. , Zeng, W. , Honaramooz, A. , Gomendio, M. , Roldan, E. R. , and Dobrinski, I. (2008). Xenografting of adult mammalian testis tissue. Anim. Reprod. Sci. 106, 65–76.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Hewitson, L. , Martinovich, C. , Simerly, C. , Takahashi, D. , and Schatten, G. (2002). Rhesus offspring produced by intracytoplasmic injection of testicular sperm and elongated spermatids. Fertil. Steril. 77, 794–801.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Honaramooz, A. , Snedaker, A. , Boiani, M. , Schöler, H. , Dobrinski, I. , and Schlatt, S. (2002). Sperm from neonatal mammalian testes grafted in mice. Nature 418, 778–781.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Honaramooz, A. , Li, M. W. , Penedo, C. T. , Meyers, S. A. , and Dobrinski, I. (2004). Accelerated maturation of primate testis by xenografting into mice. Biol. Reprod. 70, 1500–1503.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Honaramooz, A. , Megee, S. O. , Rathi, R. , and Dobrinski, I. (2007). Building a testis: formation of functional testis tissue after transplantation of isolated porcine (Sus scrofa) testis cells. Biol. Reprod. 76, 43–47.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Kim, N.-H. , Lee, J. W. , Jun, S. H. , Lee, H. T. , and Chung, K. S. (1998). Fertilization of porcine oocytes following intracytoplasmic spermatozoon or isolated sperm head injection. Mol. Reprod. Dev. 51, 436–444.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Oatley, J. M. , Reeves, J. J. , and McLean, D. J. (2005). Establishment of spermatogenesis in neonatal bovine testicular tissue following ectopic xenografting varies with donor age. Biol. Reprod. 72, 358–364.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Peters, R. M. , and Wells, K. D. (1993). Culture of pig embryos. J. Reprod. Fertil. 48((Suppl.)), 61–73.
Rathi, R. , Honaramooz, A. , Zeng, W. , Schlatt, S. , and Dobrinski, I. (2005). Germ cell fate in bovine testis tissue xenografts. Reproduction 130, 923–929.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Rathi, R. , Honaramooz, A. , Zeng, W. , Turner, R. , and Dobrinski, I. (2006). Germ cell development in equine testis tissue xenografted into mice. Reproduction 131, 1091–1098.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schlatt, S. , Kim, S. S. , and Gosden, R. (2002). Spermatogenesis and steroidogenesis in mouse, hamster and monkey testicular tissue after cryopreservation and heterotopic grafting to castrated hosts. Reproduction 124, 339–346.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schlatt, S. , Honaramooz, A. , Boiani, M. , Schöler, H. R. , and Dobrinski, I. (2003). Progeny from sperm obtained after ectopic grafting of neonatal mouse testes. Biol. Reprod. 68, 2331–2335.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Schlatt, S. , Honaramooz, A. , Ehmcke, J. , Goebell, P. J. , Rübben, H. , Dhir, R. , Dobrinski, I. , and Patrizio, P. (2006). Limited survival of adult human testicular tissue as ectopic xenograft. Hum. Reprod. 21, 384–389.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
Snedaker, A. , Honaramooz, A. , and Dobrinski, I. (2004). A game of cat and mouse: xenografting of testis tissue from domestic kittens results in complete cat spermatogenesis in a mouse host. J. Androl. 25, 926–930.
| PubMed |
Yang, H. Y. , Cox, S.-L. , Jenkin, G. , Findlay, J. , Trounson, A. , and Shaw, J. (2006). Graft site and gonadotrophin stimulation infulences the number and quality of oocytes from murine ovarian tissue grafts. Reproduction 131, 851–859.
| Crossref | GoogleScholarGoogle Scholar | PubMed |
