Cryopreservation and in vitro fertilization at the zebrafish international resource center.
Cryopreservation and in vitro fertilization at the zebrafish international resource center.
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DOI：
10.1007/978-1-60327-977-2_4
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Varga, Zoltan M
中科院分区：
文献类型：
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作者：
Carmichael, Carrie;Westerfield, Monte;Varga, Zoltan M
近几十年来，世界各地的实验室培育了数千条突变、转基因和野生型斑马鱼品系，而且还在继续生产更多的品系。与此同时，在开发可靠、高通量、标准化、长期低温保存存储方法方面花费的努力相对较少，尽管实验室和整个研究界都在努力保持大量线路的存活。维护这些宝贵遗传资源的安全可靠的方法对未来的生物医学研究至关重要。超低温保存是大规模、长期保存重要遗传物质最有效的方法。它延长了个体鱼类繁殖后代的时间，减少了维持活种群的需要，并可以防止不可替代的研究线路的灾难性损失。冷冻保存也是维持遗传资源最具成本效益的替代方案，因为它降低了动物和设施维护、人员和空间的成本。此外，它还为使用大量线条开发新型研究提供了新的机会。例如，几种遗传策略，如耕作或增强子和基因捕获，依赖于使用超低温保存来绕过几代活的有机体，直到一个菌株复活进行研究。在本章中，我们对斑马鱼国际资源中心目前使用的冷冻保存方法进行了描述和讨论。这种方法源于20多年前为斑马鱼开发的最初方案，最近得到了改进。
In recent decades, laboratories throughout the world generated several thousand mutant, transgenic, and wild-type zebrafish lines and more lines continue to be produced. At the same time, relatively little effort has been expended to develop reliable, high-throughput, standardized, long-term cryopreservation storage methods, even though laboratories and the research community as a whole struggle to maintain the large number of lines alive. Safe and reliable methods for maintaining these valuable genetic resources are vital for future biomedical research. Cryopreservation is the most efficient method for large-scale, long-term storage of important genetic materials. It extends the time offspring can be produced from individual fish, reduces the need to maintain live populations, and can prevent catastrophic loss of irreplaceable research lines. Cryopreservation is also the most cost-effective alternative for maintaining genetic resources because it reduces costs for animal and facility maintenance, personnel, and space. In addition, it provides novel opportunities to develop new types of research using large numbers of lines. For example, several genetic strategies, such as TILLING or enhancer and gene trapping, depend on the use of cryopreservation to bypass generations of live organisms until a strain is revived for research. In this chapter, we describe and discuss the current cryopreservation method used at the Zebrafish International Resource Center. This method is derived from the initial protocol developed for zebrafish over 20 years ago that has recently been refined.