In vitro male germ cell cultures of zebrafish
In vitro male germ cell cultures of zebrafish
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DOI：
10.1016/j.ymeth.2005.12.008
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发表时间：
2006-07-01
期刊：
影响因子：
4.8
通讯作者：
Sakai, Noriyoshi
中科院分区：
文献类型：
--
作者：
Sakai, Noriyoshi
受精前精子的转基因修饰具有生产转基因动物的时间轴短得多的优点。一种使用斑马鱼雄性生殖细胞的原代培养物的培养系统，其中精原细胞分化为功能性精子可以在体外发生，允许我们将外源DNA引入培养的精子中并从精子产生转基因。本章描述了雄性生殖细胞和支持细胞饲养层的共培养方法，以及用逆转录病毒引入外源DNA的方法。这种雄性生殖细胞培养系统不仅可用于生产转基因精子，而且可用于分析支持细胞对脊椎动物精子发生的调节功能。(c)2006爱思唯尔公司All rights reserved.
Transgenic modification of sperm before fertilization has the advantages of a much shorter timeline for the production of transgenic animals. A culture system using primary cultures of zebrafish male germ cells, in which the differentiation of spermatogonia to functional sperm can occur in vitro, allows us to introduce foreign DNA into the cultured sperm and to produce transgenics from the sperm. This chapter describes methods for the co-culture of male germ cells and a Sertoli cell feeder layer and the introduction of foreign DNA with retroviruses. This male germ cell culture system should prove useful not only in producing genetically modified sperm, but also in analyzing the regulatory function of Sertoli cells for spermatogenesis in vertebrates. (c) 2006 Elsevier Inc. All rights reserved.