Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control
Incorporation of bovine serum-derived albumin formulation (AlbuMAX) into a basic culture medium, MEMα, enables the completion of in vitro spermatogenesis through testicular tissue culture in mice. However, this medium was not effective in other animals. Therefore, we sought an alternative approach f...
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| Format: | Article |
| Language: | English |
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The Society for Reproduction and Development
2023-12-01
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| Series: | The Journal of Reproduction and Development |
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| Online Access: | https://www.jstage.jst.go.jp/article/jrd/70/1/70_2023-093/_pdf/-char/en |
| _version_ | 1797266483151110144 |
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| author | Takehiko OGAWA Takafumi MATSUMURA Tatsuma YAO Hiroshi KIMURA Kiyoshi HASHIMOTO Yu ISHIKAWA-YAMAUCHI Takuya SATO |
| author_facet | Takehiko OGAWA Takafumi MATSUMURA Tatsuma YAO Hiroshi KIMURA Kiyoshi HASHIMOTO Yu ISHIKAWA-YAMAUCHI Takuya SATO |
| author_sort | Takehiko OGAWA |
| collection | DOAJ |
| description | Incorporation of bovine serum-derived albumin formulation (AlbuMAX) into a basic culture medium, MEMα, enables the completion of in vitro spermatogenesis through testicular tissue culture in mice. However, this medium was not effective in other animals. Therefore, we sought an alternative approach for in vitro spermatogenesis using a synthetic medium without AlbuMAX and aimed to identify its essential components. In addition to factors known to be important for spermatogenesis, such as retinoic acid and reproductive hormones, we found that antioxidants (vitamin E, vitamin C, and glutathione) and lysophospholipids are vital for in vitro spermatogenesis. Moreover, based on our experience with microfluidic devices (MFD), we developed an alternative approach, the PDMS-ceiling method (PC method), which involves simply covering the tissue with a flat chip made of PDMS, a silicone resin material used in MFD. The PC method, while straightforward, integrates the advantages of MFD, enabling improved and uniform oxygen and nutrient supply via tissue flattening. Furthermore, our studies underscored the significance of lowering the oxygen concentration to 10–15%. Using an integrated cultivation method based on these findings, we successfully achieved in vitro spermatogenesis in rats, which has been a long-standing challenge. Further improvements in culture conditions would pave the way for spermatogenesis completion in diverse animal species. |
| first_indexed | 2024-04-25T01:01:24Z |
| format | Article |
| id | doaj.art-3934235b85474ecd9faf744d9bf2c0a3 |
| institution | Directory Open Access Journal |
| issn | 0916-8818 1348-4400 |
| language | English |
| last_indexed | 2024-04-25T01:01:24Z |
| publishDate | 2023-12-01 |
| publisher | The Society for Reproduction and Development |
| record_format | Article |
| series | The Journal of Reproduction and Development |
| spelling | doaj.art-3934235b85474ecd9faf744d9bf2c0a32024-03-11T00:27:45ZengThe Society for Reproduction and DevelopmentThe Journal of Reproduction and Development0916-88181348-44002023-12-017011910.1262/jrd.2023-093jrdImprovements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space controlTakehiko OGAWA0Takafumi MATSUMURA1Tatsuma YAO2Hiroshi KIMURA3Kiyoshi HASHIMOTO4Yu ISHIKAWA-YAMAUCHI5Takuya SATO6Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, JapanDepartment of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, JapanResearch and Development Center, Fuso Pharmaceutical Industries, Ltd., Osaka 536-8523, JapanMicro/Nano Technology Center, Tokai University, Hiratsuka 259-1292, JapanDepartment of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, JapanDepartment of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, JapanDepartment of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama 236-0004, JapanIncorporation of bovine serum-derived albumin formulation (AlbuMAX) into a basic culture medium, MEMα, enables the completion of in vitro spermatogenesis through testicular tissue culture in mice. However, this medium was not effective in other animals. Therefore, we sought an alternative approach for in vitro spermatogenesis using a synthetic medium without AlbuMAX and aimed to identify its essential components. In addition to factors known to be important for spermatogenesis, such as retinoic acid and reproductive hormones, we found that antioxidants (vitamin E, vitamin C, and glutathione) and lysophospholipids are vital for in vitro spermatogenesis. Moreover, based on our experience with microfluidic devices (MFD), we developed an alternative approach, the PDMS-ceiling method (PC method), which involves simply covering the tissue with a flat chip made of PDMS, a silicone resin material used in MFD. The PC method, while straightforward, integrates the advantages of MFD, enabling improved and uniform oxygen and nutrient supply via tissue flattening. Furthermore, our studies underscored the significance of lowering the oxygen concentration to 10–15%. Using an integrated cultivation method based on these findings, we successfully achieved in vitro spermatogenesis in rats, which has been a long-standing challenge. Further improvements in culture conditions would pave the way for spermatogenesis completion in diverse animal species.https://www.jstage.jst.go.jp/article/jrd/70/1/70_2023-093/_pdf/-char/enantioxidantsin vitro spermatogenesisorgan cultureoxygen concentrationpolydimethylsiloxane (pdms) |
| spellingShingle | Takehiko OGAWA Takafumi MATSUMURA Tatsuma YAO Hiroshi KIMURA Kiyoshi HASHIMOTO Yu ISHIKAWA-YAMAUCHI Takuya SATO Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control The Journal of Reproduction and Development antioxidants in vitro spermatogenesis organ culture oxygen concentration polydimethylsiloxane (pdms) |
| title | Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control |
| title_full | Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control |
| title_fullStr | Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control |
| title_full_unstemmed | Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control |
| title_short | Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control |
| title_sort | improvements in in vitro spermatogenesis oxygen concentration antioxidants tissue form design and space control |
| topic | antioxidants in vitro spermatogenesis organ culture oxygen concentration polydimethylsiloxane (pdms) |
| url | https://www.jstage.jst.go.jp/article/jrd/70/1/70_2023-093/_pdf/-char/en |
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