Efficient production of immunodeficient non-obese diabetic/Shi-scid IL2rγ^null mice via the superovulation technique using inhibin antiserum and gonadotropin

Abstract

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 IL2rγnull (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.

Archiving genetically altered animals: a review of cryopreservation and recovery methods for genome edited animals

“…More recent papers, however, show its effectiveness across a range of strains, and its potential for rescuing difficult models should not be overlooked. 44,45…”

Cryopreservation of mouse resources

“…The yield of ovulated oocytes using the IASe treatment was different between inbred mice (A/J: 24.9 oocytes/ female; BALB/cByJ: 90.3 oocytes/female; C3HeJ: 52.0 oocytes/female; DBA/2 J: 68.8 oocytes/female; and FVB/NJ: 25.6 oocytes/female) and outbred mice (CD1: 33.7 oocytes/ female) [44]. Among the highly immunosuppressed mouse (nonobese diabetic/Shi-scid IL2rγnull mouse), female mice aged 12 weeks produced the largest number of oocytes (70.0 oocytes/female) [45]. Therefore, the optimal age of female mice to induce ultrasuperovulation using IASe treatment also depends on the mouse strain.…”