Laboratory of Regenerative and Movement Biology, Department of Health Sciences and Technology, ETH Zurich, Schwerzenbach 8603, Switzerland.
Laboratory of Regenerative and Movement Biology, Department of Health Sciences and Technology, ETH Zurich, Schwerzenbach 8603, Switzerland; Functional Genomics Center Zurich, ETH Zurich and University of Zurich, Zurich 8057, Switzerland.
Stem Cell Reports. 2022 Sep 13;17(9):1942-1958. doi: 10.1016/j.stemcr.2022.07.005. Epub 2022 Aug 4.
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.
囊胚互补性是一种技术，旨在通过将多能干细胞 (PSCs) 注射到遗传受损的囊胚期胚胎中，在动物嵌合体中产生器官、组织或细胞类型。在这里，我们报告了在将小鼠或大鼠 PSCs 注射到携带 Tsc22d3 突变的小鼠囊胚中后，成功地对成年嵌合体中的雄性生殖细胞进行了互补，Tsc22d3 是精子发生所必需的基因。将小鼠 PSCs 注射到 Tsc22d3-Knockout (KO) 囊胚中，产生了仅包含 PSC 衍生的功能性精子的同种嵌合体。此外，将大鼠胚胎干细胞 (rESCs) 注射到 Tsc22d3-KO 胚胎中，产生了仅携带大鼠精原细胞和能够使卵母细胞受精的精子的种间小鼠-大鼠嵌合体。此外，使用单细胞 RNA 测序，我们对发生在小鼠-大鼠嵌合体睾丸中的大鼠精子发生进行了解构。总的来说，这项研究详细介绍了一种在体内进行异种生殖细胞产生的方法，这可能对大鼠转基因或濒危动物物种保护工作产生影响。
