Long Evans rat spermatogonial lines are effective germline vectors for transgenic rat production

Abstract

Long Evans rat strains are applied as research models in a broad spectrum of biomedical fields (>15,800 citations, NCBI PubMed). Here, we report an approach to genetically modify the Long Evans rat germline in donor spermatogonial stem cells. Long Evans rat spermatogonial lines were derived from freshly isolated laminin-binding spermatogonia. Laminin-binding spermatogonia were cultured over multiple passages on fibroblast feeder layers in serum-free culture medium containing GDNF and FGF2. Long Evans rat spermatogonial lines were genetically modified by transposon transduction to express a germline, tdTomato reporter gene. Donor rat spermatogonial lines robustly regenerated spermatogenesis after transplantation into testes of busulfan-treated, allogenic, Long Evans rats. Donor-derived spermatogenesis largely restored testis size in the chemically sterilized, recipient Long Evans rats. Recipient Long Evans rats stably transmitted the tdTomato germline marker to subsequent generations. Overall, Long Evans rat spermatogonial lines provided effective donor germline vectors for genetically modifying Long Evans rats.

The nature and dynamics of spermatogonial stem cells

“…Although the cultures work fine and enable one to propagate SSCs from many animals, for example mouse, human, rat and cattle (Aponte et al, 2008;Hamra et al, 2017;Oatley et al, 2016;Sadri-Ardekani et al, 2009), as yet it has not been possible to achieve complete spermatogenesisfrom SSCs up to spermatozoain vitro. However, this was possible using organ cultures of explanted immature mouse testes (Sato et al, 2011a,b).…”

Livestock Genetic Improvements through Spermatogonial Stem Cell Transplantation Technology

“…Transplantasi sel punca spermatogonia pertama kali dilakukan pada tikus untuk mengetahui proses perkembangan sel germinal pada mamalia jantan hingga berlangsungnya proses spermatogenesis (Brinster & Zimmermann 1994); (Kanatsu-Shinohara et al 2016) atau untuk memproduksi hewan transgenik (Li et al 2016, Hamra et al 2017. Setelah beberapa tahun transplantasi sel punca spermatogonia pada tikus berkembang, mulai dilakukan transplantasi sel punca spermatogonia pada mamalia besar seperti domba (Herrid et al 2009), sapi (Hill & Dobrinski 2005), babi (Zeng et al 2013), anjing (Harkey et al 2013), kera (Hermann et al 2012), dan kambing (Ciccarelli et al 2020).…”