精原干细胞（Spermatogonial stem cells, SSCs）移植受体制备，对充分挖掘种公畜繁殖力意义重大，但常用白消安腹腔注射法的高死亡率至今无法解决。避免受体死亡，提高效率，是SSCs受体制备技术研发的关键。本团队前期通过睾丸注射成功制备SSCs移植受体，获得了供体后代，避免了受体死亡，并发现SSCs消亡与恢复伴随了支持细胞（Sertoli cells, SCs）消亡与恢复，结合前人研究发现，SCs与SSCs的调控关系影响了SSCs消亡与恢复。为解决SCs与SSCs和系列生精细胞的白消安毒性调控关系科学问题，申请人拟通过小鼠和猪SSCs、SCs单独与共培养及曲细精管培养，分不同时间点检测分析白消安处理后细胞凋亡、增殖、分化调控通路上下游关键基因，探明白消安毒性作用机制与路径，突破受体制备技术效率与安全瓶颈，促进顶级公畜繁殖潜力充分挖掘和良种家畜快速扩繁。

The technology of receipient preparation for spermatogonial stem cells (SSCs) plays an important role in improving the fecundity of male animals. However, the high mortality rate caused by the common method of intraperitoneal injection of busulfan has not been solved yet. The key of SSCs receptor preparation is to improve the preparation efficiency and avoid the death of recipient. In our previous study, we successfully prepared recipients for SSCs transplantation by testicular injection, and obtained donor offspring and avoid the death of the recipient. It was found that the exhaustion and recovery of SSCs synchronized with that of sertoli cells (SCs) and the regulation of SCs and SSCs affected the exhaustion and recovery of SSCs. To investigate the regulation relationship among SCs, SSCs and series spermatogenic cells to overcome the toxicity of busulfan, we will carry out the separate culture and co-culture of SSCs and SCs and culture of seminiferous tubules from mouse and porcine. The cultured cells and the seminiferous tubules are treated with different dosages of busulfan. The key genes upstream and downstream of regulatory pathway of the apoptosis, proliferation and differentiation will be checked to explore the mechanism and path of toxicity of busulfan. The findings of this research will help to break through the bottleneck of efficiency and safety of receptor preparation technology for SSCs transplantation and promote rapid expansion of excellent breeding livestock by improving the fecundity of major male animals.