Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro
Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro
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DOI：
10.1073/pnas.0308668101
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发表时间：
2004-05-25
影响因子：
11.1
通讯作者：
Gui, JF
中科院分区：
文献类型：
--
作者：
Hong, YH;Liu, TM;Gui, JF
精原细胞是不断为下一代产生精子的男性生殖干细胞。精子发生是一个复杂的过程，经历了干细胞更新和分化的有丝分裂期、减数分裂期和精子发生的减数分裂后期。在体外完全再现精子发生是不可能的，因为在没有永生化的情况下产生正常的精原干细胞系，并且这些细胞在长期培养后无法产生可运动的精子。在这里，我们报告了一个正常的精原细胞系的衍生从一个成熟的medakfish睾丸不不朽。经过2年的140代培养，该细胞系保持稳定但依赖生长因子的增殖，二倍体核型，以及精子干细胞的表型和基因表达模式。此外，我们发现该细胞系可以进行减数分裂和精子发生以产生活动精子。因此，在培养中持续增殖和产生精子的能力是medaka精原干细胞的固有特性，而获得男性生殖细胞培养物显然不需要永生化。我们的发现和细胞系将提供一个独特的机会来研究和概括体外精子发生，并开发生殖系传播的方法。
Spermatogonia are the male germ stem cells that continuously produce sperm for the next generation. Spermatogenesis is a complicated process that proceeds through mitotic phase of stem cell renewal and differentiation, meiotic phase, and postmeiotic phase of spermiogenesis. Full recapitulation of spermatogenesis in vitro has been impossible, as generation of normal spermatogonial stem cell lines without immortalization and production of motile sperm from these cells after long-term culture have not been achieved. Here we report the derivation of a normal spermatogonial cell line from a mature medakafish testis without immortalization. After 140 passages during 2 years of culture, this cell line retains stable but growth factor-dependent proliferation, a diploid karyotype, and the phenotype and gene expression pattern of spermatogonial stem cells. Furthermore, we show that this cell line can undergo meiosis and spermiogenesis to generate motile sperm. Therefore, the ability of continuous proliferation and sperm production in culture is an intrinsic property of medaka spermatogonial stem cells, and immortalization apparently is not necessary to derive male germ cell cultures. Our findings and cell line will offer a unique opportunity to study and recapitulate spermatogenesis in vitro and to develop approaches for germ-line transmission.