In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media.
In vitro maintenance of spermatogenesis in Xenopus laevis testis explants cultured in serum-free media.
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在无血清培养基中培养的非洲爪蟾睾丸外植体中精子发生的体外维持。
DOI：
10.1095/biolreprod36.4.985
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发表时间：
1987
影响因子：
3.6
通讯作者：
Bumcrot,DA
中科院分区：
文献类型：
--
作者：
Risley,MS;Miller,A;Bumcrot,DA
在补充有牛血清白蛋白、胰岛素、转铁蛋白、卵泡刺激素、二氢睾酮、睾酮、视黄醇、抗坏血酸和生育酚的无血清培养基中培养的非洲爪蟾睾丸外植体的精子发生已维持较长时间。睾丸碎片的组织维持28天，并且在整个培养期间存在所有发育阶段。 3H-胸苷标记的次级（B型）精原细胞在28天内发育成顶体囊泡阶段的精子细胞，而标记的合子精母细胞在28天内发育成成熟的精子细胞。精原细胞增殖也在体外持续28天。生殖细胞分化不依赖于外源睾酮、抗坏血酸或生育酚，因为在缺乏这些补充剂的培养基中培养 35 天，3H 标记的精原细胞在睾丸中变成成熟的精细胞。放射自显影表明，培养 10 天的外植体中 55% 的管腔精子已在体外分化。培养10-35天的睾丸精子在活力和生育力方面与新鲜解剖的睾丸精子相似，并且与外植体培养物的精子受精的卵子正常发育成游泳蝌蚪。结果证明了在培养物中维持脊椎动物精子发生的可行性，并表明使用特定培养基对非洲爪蟾精子发生进行体外分析可能为精子发生调节机制的进化提供重要见解。
Spermatogenesis has been maintained for extended periods in Xenopus laevis testis explants cultured in serum free media supplemented with bovine serum albumin, insulin, transferrin, follicle-stimulating hormone, dihydrotestosterone, testosterone, retinol, ascorbate, and tocopherol. The organization of the testis fragmeats was maintained for 28 days, and all stages of development were present throughout the culture period. 3H-Thymidine-labeled secondary (Type B) spermatogonia developed in 28 days into spermatids at the acrosomal vesicle stage whereas labeled zygotene spermatocytes became mature spermatids in 28 days. Spermatogonial proliferation also continued in vitro for 28 days. Germ cell differentiation was not dependent upon exogenous testosterone, ascorbate, or tocopherol since 3H-labeled spermatogonia became mature spermatids in testes cultured 35 days in media lacking these supplements. Autoradiography demonstrated that 55% of the luminal sperm present in explants cultured 10 days had differentiated in vitro. Sperm from testes cultured 10-35 days were similar to sperm from freshly dissected testes with regard to motility and fecundity, and eggs fertilized with sperm from explant cultures developed normally into swimming tadpoles. The results demonstrate the feasibility of maintaining vertebrate spermatogenesis in culture and suggest that in vitro analysis of Xenopus spermatogenesis using defined media may provide important insights into the evolution of regulatory mechanisms in spermatogenesis.