全胚培养系统在啮齿动物胚胎脑中的电穿孔研究

Q2 Neuroscience

Current Protocols in Neuroscience Pub Date : 2017-01-01 DOI:10.1002/cpns.21

Takako Kikkawa, Masanori Takahashi, N. Osumi

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引用次数: 1

摘要

本单元介绍了使用胚胎日10.5天的小鼠胚胎进行哺乳动物全胚胎培养(WEC)的基本方法，包括制备高质量的立即离心(IC)大鼠血清，该血清通常用于WEC，对于体外培养的小鼠和大鼠胚胎的正常生长和发育至关重要。还介绍了不同阶段啮齿动物胚胎的替代方案。由于WEC胚胎是从子宫中剥离出来并在显微镜下操作的，因此可以克服在子宫内电穿孔所遇到的许多基因传递困难。描述了一种基因转移方法，以高度区域特异性的方式标记皮层原基的神经干/祖细胞。©2017 by John Wiley & Sons, Inc。

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Electroporation in the Rodent Embryonic Brain Using Whole Embryo Culture System

This unit describes basic methods for mammalian whole embryo culture (WEC) using embryonic day 10.5 mouse embryos, including the preparation of high‐quality immediately centrifuged (IC) rat serum that is commonly used for WEC and is essential for normal growth and development of cultured mouse and rat embryos in vitro. An alternative protocol for different stages of rodent embryos is also introduced. Since embryos for WEC are dissected out of the uterus and manipulated under the microscope, one can overcome many of the difficulties of gene delivery encountered using in utero electroporation. A description for a gene transfer method to label neural stem/progenitor cells of the cortical primordium in a highly region‐specific manner is also included. © 2017 by John Wiley & Sons, Inc.

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Current Protocols in Neuroscience Neuroscience-Neuroscience (all)

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期刊介绍： Current Protocols in Neuroscience is a one-stop resource for finding and adapting the best models and methods for all types of neuroscience experiments. Updated every three months in all formats, CPNS is constantly evolving to keep pace with the very latest discoveries and developments. A year of these quarterly updates is included in the initial CPNS purchase price.