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Rat in vitro spermatogenesis promoted by chemical supplementations and oxygen-tension control
Takafumi Matsumura1, Takuya Sato1, Takeru Abe1
1Laboratory of Biopharmaceutical and Regenerative Sciences, Institute of Molecular Medicine and Life Science, Yokohama City University Association of Medical Science, Yokohama, Kanagawa, Japan.
Abstract:
In vitro spermatogenesis (IVS) using air-liquid interphase organ culture method is possible with mouse testis tissues. The same method, however, has been hardly applicable to animals other than mice, only producing no or limited progression of spermatogenesis. In the present study, we challenged IVS of rats with modifications of culture medium, by supplementing chemical substances, including hormones, antioxidants, and lysophospholipids. In addition, reducing oxygen tension by placing tissues in an incubator of lower oxygen concentration and/or applying silicone cover ceiling on top of the tissue were effective for improving the spermatogenic efficiency. Through these modifications of the culture condition, rat spermatogenesis up to round spermatids was maintained over 70 days in the cultured tissue. Present results demonstrated a significant progress in rat IVS, revealing conditions commonly favorable for mice and rats as well as finding rat-specific optimizations. This is an important step towards successful IVS in many animal species, including humans.
Insights
Researchers optimized in vitro spermatogenesis (IVS) for rats by modifying culture media and oxygen levels. This breakthrough enables sustained rat sperm development for over 70 days, advancing IVS applications in diverse species.
Area of Science:
- Reproductive Biology
- Developmental Biology
- Organogenesis
Background:
- In vitro spermatogenesis (IVS) is established in mice using air-liquid interphase organ culture.
- Applying IVS to other animal species, like rats, has yielded limited success, with minimal or no progression of spermatogenesis.
- Developing effective IVS protocols for diverse species is crucial for reproductive research and conservation.
Purpose of the Study:
- To establish and optimize in vitro spermatogenesis (IVS) for rat testis tissues.
- To identify specific culture conditions that enhance spermatogenic efficiency in rats.
- To determine if optimized conditions could be commonly applicable to both mice and rats.
Main Methods:
- Utilized air-liquid interphase organ culture for rat testis tissues.
- Modified culture medium by supplementing hormones, antioxidants, and lysophospholipids.
- Reduced oxygen tension through lower incubator oxygen concentration and silicone cover application.
Main Results:
- Rat spermatogenesis was successfully maintained up to the round spermatid stage for over 70 days in cultured tissues.
- Optimized culture conditions, including medium modifications and reduced oxygen tension, significantly improved spermatogenic efficiency.
- Identified culture parameters that are commonly favorable for both mouse and rat IVS, alongside rat-specific optimizations.
Conclusions:
- Significant progress was made in establishing robust rat in vitro spermatogenesis (IVS).
- The study identified key factors for successful rat IVS, including specific medium additives and controlled oxygen levels.
- These findings represent a critical advancement towards achieving successful IVS in a wider range of animal species, potentially including humans.
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