Improvements in in vitro spermatogenesis: oxygen concentration, antioxidants, tissue-form design, and space control

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-12-23 DOI:10.1262/jrd.2023-093

Takehiko OGAWA, Takafumi MATSUMURA, Tatsuma YAO, Hiroshi KIMURA, Kiyoshi HASHIMOTO, Yu ISHIKAWA-YAMAUCHI, Takuya SATO

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Abstract

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.

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体外精子发生的改进：氧气浓度、抗氧化剂、组织形式设计和空间控制

在基本培养基 MEMα 中加入牛血清衍生白蛋白配方（AlbuMAX），可通过小鼠睾丸组织培养完成体外精子发生。然而，这种培养基对其他动物无效。因此，我们使用不含 AlbuMAX 的合成培养基寻找体外精子发生的替代方法，并旨在确定其基本成分。除了维甲酸和生殖激素等已知对精子发生重要的因素外，我们还发现抗氧化剂（维生素 E、维生素 C 和谷胱甘肽）和溶血磷脂对体外精子发生至关重要。此外，根据我们在微流控装置（MFD）方面的经验，我们开发了另一种方法，即 PDMS-天花板法（PC 法），该方法只需在组织上覆盖一个由 PDMS（一种用于 MFD 的硅树脂材料）制成的平面芯片。PC 方法虽然简单，但综合了中频炉的优点，通过组织扁平化实现了更好、更均匀的氧气和营养供应。此外，我们的研究还强调了将氧气浓度降至 10-15% 的重要性。在这些研究成果的基础上，我们采用综合培养方法，成功实现了大鼠的体外精子发生，这是一项长期存在的挑战。进一步改善培养条件将为在不同动物物种中完成精子发生铺平道路。图文摘要全尺寸图片

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.