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Microsurgical and Percutaneous Epididymal Sperm Aspiration for Sperm Collection from Live Mice
Auke Boersma1, Olga Olszanska1, Ingrid Walter2
1Institute of Laboratory Animal Science and Biomodels, University of Veterinary Medicine Vienna, Vienna, Austria.
Abstract:
Spermatozoa for in vitro fertilization of mouse oocytes and other methods of assisted reproduction typically are collected from the cauda epididymis of euthanized male mice. As an alternative to this terminal protocol, we developed and examined 2 methods for collecting sperm from anesthetized male mice without decreasing subsequent fertility: microsurgical epididymal sperm aspiration and, as a refinement, percutaneous epididymal sperm aspiration. Collected sperm was evaluated in terms of motility, concentration and in vitro fertilization ability. After recovery, both treated and untreated control male mice underwent in vivo fertility testing and subsequent histologic analysis of the treated male reproductive tract (epididymis and testis). In vitro fertilization using sperm recovered by the 2 collection methods was successfully achieved in all cases. The in vivo fertility test and the histologic analysis revealed no impairment of fertility and no permanent histologic alteration in the treated mice. Therefore, we recommend both techniques as simple and effective methods for recovering high-quality epididymal mouse sperm without having to euthanize fertile male mice.
Insights
Researchers developed two sperm collection methods for mice, microsurgical and percutaneous epididymal sperm aspiration, avoiding euthanasia. These techniques yield viable sperm for in vitro fertilization and maintain male fertility, offering a humane alternative for assisted reproduction research.
Area of Science:
- Reproductive Biology
- Assisted Reproductive Technologies
- Animal Models in Research
Background:
- Conventional sperm collection for mouse in vitro fertilization (IVF) requires euthanasia of male mice.
- This terminal procedure limits the availability of valuable breeding males for ongoing research.
- Alternative, non-lethal sperm retrieval methods are needed to preserve fertility and genetic lines.
Purpose of the Study:
- To develop and evaluate two novel, non-lethal sperm collection techniques from male mice.
- To assess the quality and fertilizing capacity of sperm obtained via these methods.
- To confirm the absence of adverse effects on male fertility and reproductive tract histology post-collection.
Main Methods:
- Development of microsurgical epididymal sperm aspiration (MESA).
- Refinement to percutaneous epididymal sperm aspiration (PESA).
- Evaluation of sperm motility, concentration, and in vitro fertilization (IVF) ability; in vivo fertility testing and histological analysis of reproductive tissues.
Main Results:
- Successful in vitro fertilization was achieved using sperm collected by both MESA and PESA.
- In vivo fertility tests demonstrated no impairment in the reproductive capacity of treated male mice.
- Histological examination revealed no permanent damage to the epididymis or testes in mice subjected to sperm aspiration.
Conclusions:
- Microsurgical and percutaneous epididymal sperm aspiration are effective and simple methods for obtaining high-quality mouse sperm.
- These techniques provide a humane alternative to euthanasia, preserving the fertility of male mice for assisted reproduction.
- The methods support the ethical use of animal models in reproductive science and IVF research.
