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早期鸡胚体外培养方法(新培养法)

Method for Culture of Early Chick Embryos ex vivo (New Culture).

作者信息

Psychoyos Delphine, Finnell Richard

机构信息

Center for Environmental and Genetic Medicine, Institute of Biosciences and Technology, Texas A&M Health Science Center, TX, USA.

出版信息

J Vis Exp. 2008 Oct 20(20):903. doi: 10.3791/903.

Abstract

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying the formation and patterning of brain, neural tube, somite and heart primordia. Applications of chick embryo culture include electroporation of DNA or RNA constructs in order to analyze gene function, grafts of growth factor coated beads such as FGFs and BMPs , as well as whole mount in situ hybridization and immunohistochemistry. This video demonstrates the different steps in chick embryo culture; First, the embryo is explanted in saline. Then, the embryo is centered on a glass ring. The membranes surrounding the embryo are lifted along the walls of the ring. The ring is then placed in a culture dish containing a pool of albumine. The culture dish is sealed and placed in a humid chamber, where the embryo is cultured for up to 24 hrs. Finally, the embryo is removed from the ring, fixed and processed for further applications. A troubleshooting guide is also presented.

摘要

鸡胚是研究早期胚胎发育的宝贵工具。其透明度、易获取性和易于操作的特点,使其成为研究脑、神经管、体节和心脏原基的形成与模式的理想工具。鸡胚培养的应用包括对DNA或RNA构建体进行电穿孔以分析基因功能、移植生长因子包被的珠子(如成纤维细胞生长因子和骨形态发生蛋白),以及全胚胎原位杂交和免疫组织化学。本视频展示了鸡胚培养的不同步骤;首先,将胚胎移出至盐水中。然后,将胚胎置于玻璃环的中心。围绕胚胎的膜沿着环的壁被提起。接着将环放入含有白蛋白池的培养皿中。培养皿密封后置于湿度箱中,胚胎在其中培养长达24小时。最后,将胚胎从环中取出,固定并进行进一步处理以用于其他应用。还提供了一份故障排除指南。

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