Objective To establish a comprehensive system for mouse strain conservation through natural mating, sperm cryopreservation and assisted reproductive technology. Methods Mouse fresh epididymal sperm were collected by vasectomy of unilateral testis and the vasectomized mice were naturally mated for breeding. If no satisfactory results obtained from natural breeding, then the cryopreserved sperm were thawed for In Vitro Fertilization (IVF). If no results obtained, Intracytoplas-mic Sperm Injection (ICSI) was used to reconstructed embryos and which were finally transferred into pseudopregnant recipient female mouse. Results The results show that parts of mice vasectomized unilateral testis such as BDF1 and C57BL/6J mice could be used for natural breeding. Among the remaining sterile males, only ICR mice could be preserved through IVF while ICSI technology was the last approach to save GFP mice with C57BL/6J background. Conclusion The genetic background of mouse sperm cryopreservation was a key factor in IVF and setup a comprehensive system, which could achieve the purpose of strain conservation while mouse strains could be utilized at maximum extent.%目的 利用活体保种、精子冷冻技术及辅助生殖技术建立一套安全的综合性保种体系.方法 以单侧睾丸摘除方法获得ICR、BDF1、C57BL/6J和GFP四种小鼠的附睾精子进行冷冻保存实验,术后恢复的公鼠继续用于自然繁育.如果无法自然繁育,则将冻融精子用于体外受精(IVF).如果冻融精子仍无法受精获得后代,则需要借助卵胞质内单精子注射技术(ICSI)进行胚胎构建,胚胎移植,直至获得后代.结果 单侧睾摘除的BDF1、C57BL/6J的公鼠可以继续繁育；不育的ICR和GFP小鼠的冻融精子IVF后,只有ICR小鼠可以得到正常的胚胎和子鼠;而以C57BL/6J 为遗传背景的GFP小鼠,其冻融精子只有采用ICSI技术,才可以得到子代.结论 精子冻融小鼠的遗传背景是影响IVF成功与否的关键性因素,同时还建立了一种以小鼠精子为主导的综合性保种体系,既可以达到品系保存的目的,同时又可以最大限度地利用小鼠品系.
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