Severe immunodeficient mice are an essential tool for the examination of the efficacy and safety of new therapeutic technologies as a humanized model. Previously, non-obese diabetic (NOD)/Shi-scid IL2rγ^null (NOG) mice were established as immunodeficient mice by combining interleukin-2 receptor-γ chain-knockout mice and NOD/Shi-scid mice. The NOG mice are used frequently in the research of therapeutic monoclonal antibodies and regenerative medicine for human diseases. Establishment of an efficient production system of NOG mice, using optimized reproductive techniques, is required to accelerate research. In this study, we investigated the efficacy of the superovulation technique using equine chorionic gonadotropin (eCG) and inhibin antiserum (IAS) in NOG mice of various ages (4, 8, 12, 24, or 54 weeks). Additionally, we examined the fertilizing and developmental ability of the oocytes through in-vitro fertilization using frozen–thawed sperm, embryo culture and embryo transfer. The results showed that NOG mice produced the highest number of oocytes at 12 weeks old following the co-administration of eCG and IAS (collectively IASe) (70 oocytes/female). IASe was more effective in increasing the number of oocytes v. eCG at all ages. The IASe-derived oocytes demonstrated the ability to fertilize and develop into blastocysts and pups. Finally, we demonstrated that three strains of genetically modified NOG mice were efficiently produced through the optimized reproductive techniques. In summary, we developed an efficient system for the production of immunodeficient mice using 12-week-old, IASe-treated female NOG mice.

严重免疫缺陷小鼠是检查新治疗技术作为人性化模型的有效性和安全性的重要工具。以前，非肥胖糖尿病 (NOD)/Shi- scid IL2rγ^无效(NOG) 小鼠通过结合白细胞介素 2 受体-γ 链敲除小鼠和 NOD/Shi-scid 小鼠被建立为免疫缺陷小鼠。NOG 小鼠经常用于治疗性单克隆抗体和人类疾病再生医学的研究。需要使用优化的繁殖技术建立有效的 NOG 小鼠生产系统，以加快研究速度。在这项研究中，我们研究了使用马绒毛膜促性腺激素 (eCG) 和抑制素抗血清 (IAS) 在不同年龄（4、8、12、24 或 54 周）的 NOG 小鼠中进行超排卵技术的功效。此外，我们通过体外检测了卵母细胞的受精和发育能力。使用冻融精子、胚胎培养和胚胎移植进行受精。结果表明，在同时施用 eCG 和 IAS（统称为 IASe）（70 个卵母细胞/雌性）后，NOG 小鼠在 12 周龄时产生的卵母细胞数量最多。IASe 在增加所有年龄段的卵母细胞与心电图的数量方面更有效。IASe 衍生的卵母细胞显示出受精和发育成囊胚和幼崽的能力。最后，我们证明了通过优化的生殖技术有效地生产了三株转基因 NOG 小鼠。总之，我们开发了一种使用 12 周龄、IASe 处理的雌性 NOG 小鼠生产免疫缺陷小鼠的有效系统。