鱼类精原干细胞的培养

圆圆脸

      来这个论坛有段时间了，学习了不少知识，看大家都做的是人类或小鼠的胚胎干细胞，发个其他物种的，鱼类方面，模式生物青鱂的生殖干细胞的培养，第一次发帖。3 o" `1 X1 o% B3 y: W5 Q+ s
Proc Natl Acad Sci U S A. 2004 May 25;101(21):8011-6. Epub 2004 May 12.& l% I* W3 ^7 g3 z
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Establishment of a normal medakafish spermatogonial cell line capable of sperm production in vitro.
! g! Q$ ]) O3 EHong Y, Liu T, Zhao H, Xu H, Wang W, Liu R, Chen T, Deng J, Gui J.
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Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260.
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6 g( h5 B+ l: o7 X2 [/ @( K" t/ ?Abstract
6 n+ W, V  c6 G: b) E# lSpermatogonia 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.$ Y7 n, |& f9 ~/ L( L
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