Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis and fertility in male animals. In agricultural perspectives, SSC transplantation holds potential in expanding the availability of gametes from elite livestock rapidly, thus offering a highly efficient reproductive technique for livestock production. A limiting factor in application of this technique is acquisition of non-endogenous-SSC recipients, especially from large domestic animals. To explore a simple method to ablate pig endogenous SSCs and prepare transplant recipients, we directly injected different doses of busulfan into the testis tissue of Duroc pigs. Following injection, no significant health problems and body weight loss were observed in pigs treated with 3 mg/kg busulfan. Morphological analysis and gene expression assays showed that busulfan treatment efficiently removed endogenous SSCs, but sustained an intact Sertoli cells structure in pig testis. Compared with 5 mg/kg busulfan treatment group, 3 mg/kg busulfan treatment resulted in a complete SSCs ablation with less toxicity, providing a suitable environment for transplantation of exogenous SSCs. This study showed that intratesticular injection of busulfan in pigs to prepare recipients was an efficient and safe method, which lays the groundwork for the application of pig SSC transplantation.

精原干细胞（SSC）是雄性动物精子发生和繁殖的基础。从农业的角度来看，南南合作移植具有迅速扩大精英牲畜配子供应的潜力，从而为牲畜生产提供了高效的繁殖技术。应用该技术的限制因素是非内源性SSC受体的获得，特别是从大型家畜中获得。为了探索一种消融猪内源性SSC并准备移植受者的简单方法，我们直接将不同剂量的白消安注射到杜洛克猪的睾丸组织中。注射后，在用3 mg / kg白消安治疗的猪中未观察到明显的健康问题和体重减轻。形态分析和基因表达分析表明，白消安治疗有效去除了内源性SSC，但在猪睾丸中维持了完整的支持细胞结构。与5 mg / kg白消安治疗组相比，3 mg / kg白消安治疗导致完全的SSC消融，且毒性较低，为外源性SSC的移植提供了合适的环境。这项研究表明，向猪睾丸内注射白消安制备容器是一种安全有效的方法，这为猪SSC移植的应用奠定了基础。为外源性SSC的移植提供合适的环境。这项研究表明，向猪睾丸内注射白消安制备容器是一种安全有效的方法，这为猪SSC移植的应用奠定了基础。为外源性SSC的移植提供合适的环境。这项研究表明，向猪睾丸内注射白消安制备容器是一种安全有效的方法，这为猪SSC移植的应用奠定了基础。