Antioxidant vitamins and lysophospholipids are critical for inducing mouse spermatogenesis under organ culture conditions
Antioxidant vitamins and lysophospholipids are critical for inducing mouse spermatogenesis under organ culture conditions
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DOI：
10.1096/fj.202000245r
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发表时间：
2020-05-31
期刊：
影响因子：
4.8
通讯作者：
Ogawa, Takehiko
中科院分区：
文献类型：
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作者：
Sanjo, Hiroyuki;Yao, Tatsuma;Ogawa, Takehiko
通过在基础培养基中仅添加经色谱纯化的牛血清白蛋白产物（AlbuMAX），可以使用经典器官培养方法进行体外小鼠精子发生，这表明AlbuMAX含有小鼠精子发生所需的所有化学因子。然而，由于这些因素的身份尚不清楚，在培养基和我们的理解精子发生所需的营养和信号物质的改进受到阻碍。在本研究中，使用不含AlbuMAX的化学成分确定的培养基（CDM）评价每种辅助因子及其组合对精子发生的诱导作用。与体内条件相似，需要维甲酸、三碘甲腺原氨酸（T-3）和睾酮（T）。基于AlbuMAX、胎牛血清和成年牛血清之间生精能力的差异，我们鉴定了α-生育酚，当与抗坏血酸和谷胱甘肽结合时，α-生育酚强烈促进精子发生。还观察到使用两种不同方法从AlbuMAX中提取的脂质诱导精子发生的能力存在差异。这导致了溶血磷脂，特别是溶血磷脂酰胆碱，溶血磷脂酸和溶血磷脂酰丝氨酸，作为精子发生的重要分子的鉴定。基于这些结果配制的新CDM与含AlbuMAX的培养基一样有效地诱导和促进精子发生。利用CDM技术进行体外精子发生的研究，为研究体内实验所不能实现的精子发生提供了一个独特的实验系统。
In vitro mouse spermatogenesis using a classical organ culture method became possible by supplementing basal culture medium with only the product of bovine serum albumin purified by chromatography (AlbuMAX), which indicated that AlbuMAX contained every chemical factor necessary for mouse spermatogenesis. However, since the identity of these factors was unclear, improvements in culture media and our understanding of the nutritional and signal substances required for spermatogenesis were hindered. In the present study, chemically defined media (CDM) without AlbuMAX was used to evaluate each supplementary factor and their combinations for the induction of spermatogenesis. Similar to in vivo conditions, retinoic acid, triiodothyronine (T-3), and testosterone (T) were needed. Based on differences in spermatogenic competence between AlbuMAX, fetal bovine serum, and adult bovine serum, we identified alpha-tocopherol, which strongly promoted spermatogenesis when combined with ascorbic acid and glutathione. Differences were also observed in the abilities of lipids extracted from AlbuMAX using two different methods to induce spermatogenesis. This led to the identification of lysophospholipids, particularly lysophosphatidylcholine, lysophosphatidic acid, and lysophosphatidylserine, as important molecules for spermatogenesis. New CDM formulated based on these results induced and promoted spermatogenesis as efficiently as AlbuMAX-containing medium. In vitro spermatogenesis with CDM may provide a unique experimental system for research on spermatogenesis that cannot be performed in in vivo experiments.