Xenotransplantation of Human Spermatogonia Into Various Mouse Recipient Models

Dongli Liang; Qi Sun; Zijue Zhu; Chuanyun Wang; Shicheng Ye; Zheng Li; Yuan Wang

doi:10.3389/fcell.2022.883314

Xenotransplantation of Human Spermatogonia Into Various Mouse Recipient Models

Front Cell Dev Biol. 2022 May 23:10:883314. doi: 10.3389/fcell.2022.883314. eCollection 2022.

Authors

Dongli Liang¹, Qi Sun², Zijue Zhu³, Chuanyun Wang², Shicheng Ye², Zheng Li³, Yuan Wang⁴

Affiliations

¹ Laboratory Animal Center, Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai, China.
² Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China.
³ Department of Andrology, The Center for Men's Health, Urologic Medical Center, Shanghai Key Laboratory of Reproductive Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
⁴ Department of Animal Sciences, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI, United States.

Abstract

Spermatogonial stem cells are the foundation of continuous spermatogenesis in adult mammals. Xenograft models have been established to define human SSCs, mostly using infertile and immune-deficient mice as the recipients for human germ cell transplantation. However, it is time-consuming to prepare such recipients using irradiation or chemotherapeutic agents, and this approach may also introduce confounding factors when residual endogenous germ cells recover in transplanted recipients. It remains to be determined whether immune-competent genetically infertile mice can be suitable recipients for xenotransplantation. In this study, we observed similar engraftment efficiencies when using spermatogonia from human biopsied testes across immune-deficient nude mice, immune-competent ICR mice, and genetically infertile Kit ^w/w-v mice, suggesting minimal immunological rejection from immune-competent mouse recipients upon xenotransplantation of human germ cells. More importantly, we derived EpCAM negative and TNAP positive spermatogonia-like cells (SLCs) from human pluripotent stem cells (PSCs), which highly expressed spermatogonial markers including PLZF, INTERGRINα6, TKTL1, CD90, and DRMT3. We found that upon transplantation, these SLCs proliferated and colonized at the basal membrane of seminiferous tubules in testes of both immune-deficient nude mice and Kit ^w/w-v mice, though complete spermatogenesis would likely require supporting human signaling factors and microenvironment. Taken together, our study functionally defined the cell identity of PSC-derived SLCs, and supported xenotransplantation using genetically infertile recipients as a convenient model for functionally evaluating spermatogonia derived from different species.

Keywords: germ cells; human pluripotent stem cells; immunecompetent mouse; spermatogonial stem cells; transplantation.