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In vivo serial sampling of epididymal sperm in mice
Gonzalo Moreno Del Val1, Patricia Muñoz Robledano
1Laboratorio de Criopreservación, Servicio de Experimentación Animal UMH, Instituto de Neurociencias de Alicante, Consejo Superior de Investigaciones Científicas/Universidad Miguel Hernández, Avenida Ramón y Cajal s/n, San Juan de Alicante, 03550 Alicante, Spain. g.moreno@umh.es
Abstract:
This study was undertaken to refine the techniques of in vivo collection of sperm in the mouse. The principal objective was to offer a viable, safe and reliable method for serial collection of in vivo epididimary sperm through the direct puncture of the epididymis. Six C57Bl/6J males were subjected to the whole experiment. First we obtain a sperm sample of the right epididymis, and perform a vasectomy on the left side. This sample was used in an in vitro fertilization (IVF) experiment while the males were individually housed for 10 days to let them recover from the surgery, and then their fertility was tested with natural matings until we obtained a litter of each one. After that, the animals were subjected another time to the same process (sampling, recover and natural mating). The results of these experiments were a fertilization average value of 56.7%, and that all the males had a litter in the first month after the natural matings. This study documented the feasibility of the epididimary puncture technique to in vivo serial sampling of sperm in the mouse.
Insights
This study presents a safe and reliable method for serial sperm collection in mice using epididymal puncture. This technique enables repeated in vivo sperm sampling for fertility studies and assisted reproduction.
Area of Science:
- Reproductive Biology
- Animal Models
- Sperm Collection Techniques
Background:
- In vivo sperm collection is crucial for reproductive research and assisted reproduction technologies.
- Existing methods for serial sperm collection in mice can be invasive or unreliable.
- There is a need for refined, safe, and efficient techniques for obtaining epididymal sperm in vivo.
Purpose of the Study:
- To develop and validate a viable, safe, and reliable method for serial in vivo collection of epididymal sperm in mice.
- To assess the feasibility of direct epididymal puncture for repeated sperm sampling.
- To evaluate the impact of the procedure on male mouse fertility and sperm quality.
Main Methods:
- Six C57Bl/6J male mice were used for the study.
- A novel technique involving direct puncture of the epididymis for sperm collection was employed.
- Procedures included initial sperm sampling, vasectomy, recovery period, in vitro fertilization (IVF) trials, and natural mating fertility tests.
Main Results:
- The epididymal puncture technique demonstrated feasibility for in vivo serial sperm sampling.
- An average fertilization rate of 56.7% was achieved in in vitro fertilization experiments.
- All male mice successfully produced litters within one month following natural mating trials after sperm collection.
Conclusions:
- The direct epididymal puncture technique is a feasible, safe, and reliable method for serial in vivo sperm collection in mice.
- This refined technique supports applications in reproductive research, sperm banking, and assisted reproductive technologies.
- The study validates the efficacy of the procedure without compromising the overall fertility of the male mice.
