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Updated: May 17, 2025
Extra Cellular Matrix-Based and Extra Cellular Matrix-Free Generation of Murine Testicular Organoids
Published on: October 7, 2020
A long-term mouse testis organ culture system to identify germ cell damage induced by chemotherapy
Satoshi Yokota1, Kiyoshi Hashimoto2,3, Takuya Sato2
1Division of Cellular and Molecular Toxicology, Center for Biological Safety and Research, National Institute of Health Sciences, Kanagawa 210-9501, Japan.
Abstract:
We previously developed the acrosin-green fluorescent protein (GFP) transgenic neonatal mouse organ culture system for rapid and accurate assessment of testicular toxicity. This system effectively evaluates drug-induced toxicity in male germ cells before meiotic entry but cannot assess post-meiotic germ cell toxicity. For many chemicals, the specific stage of germ cell differentiation that is susceptible to toxicity remains unclear, highlighting the need for new methods. In this study, we incubated neonatal mouse testis organ cultures for 35 days to allow post-meiotic cells to develop. The tissue was then exposed to cisplatin to determine the cells that are targeted and to assess the reversibility of the toxicity. We monitored changes in tissue volume and GFP fluorescence, which tracks the progression of spermatogenesis, and confirmed findings by histological analysis. Cisplatin inhibited tissue growth and reduced GFP fluorescence in a concentration-dependent manner. Higher concentrations targeted not only spermatogonia, but also spermatocytes and spermatids. Recovery from toxicity was observed at clinically relevant doses. This study demonstrates that long-term mouse testis organ culture can be used to assess testicular toxicity, enabling the identification of specific germ cell stages targeted by chemicals such as cisplatin.
Insights
Long-term mouse testis organ culture effectively assesses chemical toxicity in developing sperm cells. This method identifies specific germ cell stages targeted by drugs like cisplatin and evaluates recovery.
Area of Science:
- Reproductive Toxicology
- Developmental Biology
- Pharmacology
Background:
- Neonatal mouse organ culture models are crucial for assessing testicular toxicity.
- Current models are limited to evaluating toxicity in pre-meiotic germ cells.
- Assessing post-meiotic germ cell toxicity is essential for understanding chemical effects on male fertility.
Purpose of the Study:
- To establish and validate a long-term neonatal mouse testis organ culture system for assessing post-meiotic germ cell toxicity.
- To determine the specific germ cell stages targeted by cisplatin and evaluate the reversibility of its toxicity.
- To enhance the assessment of chemical-induced testicular toxicity.
Main Methods:
- Incubation of neonatal mouse testis organ cultures for 35 days to allow post-meiotic germ cell development.
- Exposure of cultured testes to varying concentrations of cisplatin.
- Monitoring of tissue volume and acrosin-green fluorescent protein (GFP) fluorescence to track spermatogenesis progression.
- Histological analysis to confirm observed toxicity and cellular targets.
Main Results:
- Cisplatin exposure inhibited testis tissue growth and reduced GFP fluorescence in a dose-dependent manner.
- Higher cisplatin concentrations were toxic to spermatogonia, spermatocytes, and spermatids.
- Recovery from cisplatin-induced toxicity was observed at clinically relevant doses.
Conclusions:
- Long-term mouse testis organ culture is a viable method for assessing testicular toxicity, including effects on post-meiotic germ cells.
- This model allows for the identification of specific germ cell populations susceptible to chemical agents like cisplatin.
- The system facilitates the evaluation of toxicity reversibility, providing valuable insights into male reproductive health risks.
