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Updated: Jun 1, 2026
Germ Cell Transplantation and Testis Tissue Xenografting in Mice
Published on: February 6, 2012
Efficient and safe recipient preparation for transplantation of mouse spermatogonial stem cells: pretreating testes
Wenzhi Ma1, Lei An, Zhonghong Wu
1Key Laboratory of Animal Genetics and Breeding of the Ministry of Agriculture, College of Animal Sciences and Technology, China Agricultural University, Beijing, China.
Abstract:
Recipient preparation is of prime importance for the successful transplantation of spermatogonial stem cells (SSCs). Busulfan destroys endogenous germs cells and is commonly used for recipient preparation. However, busulfan produces significant side effects, including systemic toxicity, and it is lethal in certain species. The side effects associated with busulfan compromise the efficiency of SSC transplantation and threaten the safety of recipients. Here, we show that heat shock treatment of testes can be used as an alternative to busulfan treatment. Fourteen days after heat shock treatment, mice received a testicular injection of donor germ cells expressing enhanced green fluorescent protein (EGFP). Busulfan-treated mice were used as controls. Two months after transplantation, the number (12 ± 1 mm) and length (30.46 ± 5.23 mm) of EGFP-expressing testicular colonies in heat shock-treated recipients were not significantly different from those in busulfan-treated recipients. Furthermore, healthy EGFP-expressing offspring were obtained after intracytoplasmic injection of round spermatids recovered from heat shock-treated recipients. This result indicates that donor SSCs undergo complete spermatogenesis in the heat shock-treated testes of recipients. Our findings demonstrate the feasibility of using heat shock for the preparation of recipients before SSC transplantation in mice. Heat shock may prove to be useful for recipient preparation in mammalian species in which busulfan produces significant toxicity.
Insights
Heat shock treatment offers a safer alternative to busulfan for preparing recipients for spermatogonial stem cell (SSC) transplantation. This method successfully supports complete spermatogenesis and offspring production, highlighting its potential in mammalian species.
Area of Science:
- Reproductive Biology
- Stem Cell Biology
- Toxicology
Background:
- Recipient preparation is crucial for successful spermatogonial stem cell (SSC) transplantation.
- Busulfan is a common agent for eliminating endogenous germ cells but causes significant systemic toxicity and lethality in some species.
- These adverse effects limit SSC transplantation efficiency and recipient safety.
Purpose of the Study:
- To investigate heat shock treatment as a safer alternative to busulfan for recipient preparation in SSC transplantation.
- To evaluate the efficacy of heat shock in supporting SSC colonization and spermatogenesis.
- To assess the potential for generating offspring from transplanted SSCs in heat shock-treated recipients.
Main Methods:
- Testicular heat shock was applied to mice as a recipient preparation method.
- Donor germ cells expressing enhanced green fluorescent protein (EGFP) were transplanted into heat-treated testes.
- Busulfan-treated mice served as controls for comparison.
- EGFP-expressing testicular colonies were analyzed for number and length post-transplantation.
- Round spermatids from treated recipients were used for intracytoplasmic sperm injection to assess fertility and offspring viability.
Main Results:
- Heat shock treatment resulted in EGFP-expressing testicular colonies comparable in number and length to those in busulfan-treated controls.
- Complete spermatogenesis was confirmed by the successful generation of healthy EGFP-expressing offspring.
- No significant differences were observed between heat shock and busulfan groups regarding colony formation and reproductive success.
Conclusions:
- Heat shock treatment is a feasible and effective method for preparing recipients for SSC transplantation in mice.
- This approach avoids the systemic toxicity associated with busulfan.
- Heat shock presents a promising alternative for recipient preparation in mammalian SSC transplantation, especially in species sensitive to busulfan toxicity.
