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Updated: May 24, 2026
Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit
Published on: December 12, 2011
Fertile offspring derived from mouse spermatogonial stem cells cryopreserved for more than 14 years
Xin Wu1, Shaun M Goodyear, Lara K Abramowitz
1Department of Animal Biology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Background:
Approximately 80% of childhood cancers can now be cured but a side effect of treatment results in about one-third of the surviving boys being infertile or severely subfertile when they reach reproductive age. Currently, more than 1 in 5000 men of reproductive age who are childhood cancer survivors suffer from this serious quality of life problem. It is possible to obtain a testicular biopsy before treatment to preserve the spermatogonial stem cells (SSCs) of the male by cryopreservation, but the results of long-term storage of SSCs on their subsequent functional ability to generate normal offspring has not been examined in any mammalian species. Moreover, it will be necessary to increase the number of these cryopreserved SSCs to remove any contaminating malignant cells and assure regeneration of spermatogenesis.
Methods And Results:
In this report, we demonstrate that long-term cryopreservation (>14 years) of testis cells from mouse, rat, rabbit and baboon safeguards SSC viability, and that these cells can colonize the seminiferous tubules of recipient testes. Moreover, mouse and rat SSCs can be cultured and re-establish complete spermatogenesis, and fertile mouse progeny without apparent genetic or epigenetic errors were generated by the sperm produced.
Conclusions:
These findings provide a platform for fertility preservation in prepubertal boys undergoing gonadotoxic treatments.
Insights
Long-term cryopreservation of spermatogonial stem cells (SSCs) maintains their viability and function. This research confirms that cryopreserved SSCs can restore fertility in mammals, offering hope for childhood cancer survivors.
Area of Science:
- Reproductive biology
- Stem cell science
- Oncology
Background:
- Childhood cancer treatments can cause infertility in approximately one-third of male survivors.
- Preserving spermatogonial stem cells (SSCs) via cryopreservation is a potential fertility preservation method.
- The long-term functional capacity of cryopreserved SSCs has not been previously evaluated in mammals.
Purpose of the Study:
- To assess the viability and functional capacity of long-term cryopreserved SSCs.
- To determine if cryopreserved SSCs can restore spermatogenesis and fertility.
- To establish a platform for fertility preservation in prepubertal boys.
Main Methods:
- Cryopreservation of testis cells from mouse, rat, rabbit, and baboon for over 14 years.
- Transplantation of cryopreserved SSCs into recipient testes.
- Culture of mouse and rat SSCs to assess spermatogenesis and fertility.
Main Results:
- Long-term cryopreservation (>14 years) preserved SSC viability in multiple species.
- Cryopreserved SSCs successfully colonized seminiferous tubules in recipient testes.
- Fertile mouse offspring were produced from cryopreserved and cultured SSCs without genetic or epigenetic errors.
Conclusions:
- Long-term cryopreservation of SSCs is a viable method for fertility preservation.
- This study provides a foundation for fertility preservation strategies for prepubertal boys undergoing cancer treatment.
- Successful generation of fertile offspring demonstrates the potential of this approach.
