A rapid and effective nonsurgical artificial insemination protocol using the NSET™ device for sperm transfer in mice without anesthesia

Artificial insemination (AI) is an assisted reproductive technique that is implemented successfully in humans as a fertility treatment, performed extensively for commercial breeding of livestock, and is also successful in laboratory rodents. AI in the mouse may be especially useful for breeding of transgenic or mutant mice with fertility problems, expansion of mouse colonies, and as an alternative to in vitro fertilization. Nonsurgical AI techniques for the mouse have been described previously but are not often implemented due to technical difficulties. Here we compare various protocols for preparation of CD1 recipients prior to AI for naïve (in estrus), ovulation-induced, and superovulated females. Timing of hormone administration relative to sperm delivery is also compared. An improved protocol for nonsurgical AI in mice is described, which incorporates a convenient hormone administration schedule for female recipients and rapid, non-stressful sperm transfer without the need for anesthesia or analgesia.