A novel culture system for mouse spermatid maturation which produces elongating spermatids capable of inducing calcium oscillation during fertilization and embryonic development
A novel culture system for mouse spermatid maturation which produces elongating spermatids capable of inducing calcium oscillation during fertilization and embryonic development
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DOI：
10.1007/s10815-010-9442-3
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发表时间：
2010-09-01
影响因子：
3.1
通讯作者：
Sato, Kahei
中科院分区：
文献类型：
--
作者：
Hasegawa, Hisataka;Terada, Yukihiro;Sato, Kahei
目的：建立小鼠圆形精子体外培养系统，模拟精子发生并评估卵母细胞激活能力。从B6D2F1雄性小鼠睾丸组织中分离出圆形精子和支持细胞，在睾酮和重组卵泡刺激素的作用下共培养。对体外培养的精子进行ICSI后细胞核形态和凝结、受精和发育率以及Ca2+振荡模式的检测。培养的精子在形态和核凝聚方面与正常的精子相似。新鲜和培养的伸长精子受精率和发育率无显著差异。此外，在卵母细胞激活的初始阶段，培养精子表现出与新鲜拉长精子相似的Ca2+振荡模式。这些数据表明，精子和支持细胞的共培养系统，补充睾酮和重组FSH，支持圆形精子向细长精子的正常分化，通过其形态、核凝聚和卵母细胞激活能力进行评估。
To establish an in vitro culture system for mouse round spermatids that models spermiogenesis and enables the assessment of oocyte activation ability.Round spermatids and Sertoli cells were isolated from testicular tissues of B6D2F1 male mice and co-cultured in the presence of testosterone and recombinant FSH. Cultured spermatids were examined for morphology and condensation of nuclei, fertilization and development rate, and Ca2+ oscillation pattern after ICSI.The cultured spermatids elongated and resembled normal elongating spermatids in terms of both morphology and nuclear condensation. No significant differences in fertilization and development rates were observed between fresh and cultured elongating spermatids. Moreover, cultured spermatids showed similar Ca2+ oscillation patterns to fresh elongating spermatids during an initial stage in oocyte activation.These data suggest that a co-culture system of spermatids and Sertoli cells, supplemented with testosterone and recombinant FSH, supports normal differentiation of round spermatids into elongating spermatids, as assessed by their morphology, nuclear condensation, and oocyte activation ability.