Cells (Nov 2023)

<i>DAZL</i> Knockout Pigs as Recipients for Spermatogonial Stem Cell Transplantation

  • Nathalia L. M. Lara,
  • Taylor Goldsmith,
  • Paula Rodriguez-Villamil,
  • Felipe Ongaratto,
  • Staci Solin,
  • Dennis Webster,
  • Uyanga Ganbaatar,
  • Shane Hodgson,
  • Stanislas M. A. S. Corbière,
  • Alla Bondareva,
  • Daniel F. Carlson,
  • Ina Dobrinski

DOI
https://doi.org/10.3390/cells12212582
Journal volume & issue
Vol. 12, no. 21
p. 2582

Abstract

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Spermatogonial stem cell (SSC) transplantation into the testis of a germ cell (GC)-depleted surrogate allows transmission of donor genotype via donor-derived sperm produced by the recipient. Transplantation of gene-edited SSCs provides an approach to propagate gene-edited large animal models. DAZL is a conserved RNA-binding protein important for GC development, and DAZL knockout (KO) causes defects in GC commitment and differentiation. We characterized DAZL-KO pigs as SSC transplantation recipients. While there were GCs in 1-week-old (wko) KO, complete GC depletion was observed by 10 wko. Donor GCs were transplanted into 18 DAZL-KO recipients at 10–13 wko. At sexual maturity, semen and testes were evaluated for transplantation efficiency and spermatogenesis. Approximately 22% of recipient seminiferous tubules contained GCs, including elongated spermatids and proliferating spermatogonia. The ejaculate of 89% of recipients contained sperm, exclusively from donor origin. However, sperm concentration was lower than the wild-type range. Testicular protein expression and serum hormonal levels were comparable between DAZL-KO and wild-type. Intratesticular testosterone and Leydig cell volume were increased, and Leydig cell number decreased in transplanted DAZL-KO testis compared to wild-type. In summary, DAZL-KO pigs support donor-derived spermatogenesis following SSC transplantation, but low spermatogenic efficiency currently limits their use for the production of offspring.

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