Exclusive generation of rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells
Joel Zvick, Monika Tarnowska-Sengül, Adhideb Ghosh, Nicola Bundschuh, Pjeter Gjonlleshaj, Laura C. Hinte, Christine L. Trautmann, Falko Noé, Xhem Qabrati, Seraina A. Domenig, Inseon Kim, Thomas Hennek, Ferdinand von Meyenn, Ori Bar‐Nur
Abstract
Blastocyst complementation denotes a technique that aims to generate organs, tissues, or cell types in animal chimeras via injection of pluripotent stem cells (PSCs) into genetically compromised blastocyst-stage embryos. Here, we report on successful complementation of the male germline in adult chimeras following injection of mouse or rat PSCs into mouse blastocysts carrying a mutation in Tsc22d3, an essential gene for spermatozoa production. Injection of mouse PSCs into Tsc22d3-Knockout (KO) blastocysts gave rise to intraspecies chimeras exclusively embodying PSC-derived functional spermatozoa. In addition, injection of rat embryonic stem cells (rESCs) into Tsc22d3-KO embryos produced interspecies mouse-rat chimeras solely harboring rat spermatids and spermatozoa capable of fertilizing oocytes. Furthermore, using single-cell RNA sequencing, we deconstructed rat spermatogenesis occurring in a mouse-rat chimera testis. Collectively, this study details a method for exclusive xenogeneic germ cell production in vivo, with implications that may extend to rat transgenesis, or endangered animal species conservation efforts.