Sendai virus-mediated transduction of mammalian spermatogonial stem cells†

Area of Science:

• Reproductive Biology
• Virology
• Cell Biology

Background:

• Spermatogonial stem cells (SSCs) are crucial for spermatogenesis but difficult to transfect.
• Existing viral vectors face challenges with genome integration and long-term gene expression in SSCs.

Purpose of the Study:

• To evaluate Sendai virus (SV) as a novel vector for SSC transfection.
• To assess the efficiency and longevity of gene expression mediated by SV in SSCs.
• To determine if SV transfection impacts SSC function and spermatogenesis.

Main Methods:

• Transfection of cultured germline stem (GS) cells and freshly isolated SSCs using SV.
• Spermatogonial transplantation of SV-transfected GS cells into infertile mice.
• Microinjection of SV into seminiferous tubules of immature mice.
• Xenogeneic transplantation of SV-infected cells from various species.

Main Results:

• SV efficiently transduced GS cells and freshly isolated SSCs, maintaining gene expression for over 5 months.
• SV transfection did not impede spermatogenesis progression after transplantation.
• SV successfully transduced SSCs and Sertoli cells in vivo, but not Leydig or peritubular cells.
• SV demonstrated efficacy in transducing SSCs from multiple mammalian species, including hamster, rabbit, and monkey.

Conclusions:

• Sendai virus is an effective RNA vector for transducing mammalian spermatogonial stem cells and Sertoli cells.
• SV overcomes limitations of previous vectors, offering sustained gene expression and functional recovery of SSCs.
• SV holds promise for advancing research and therapeutic applications involving SSCs.