利用全胚胎培养系统对啮齿动物胚胎脑进行电穿孔

Q2 Neuroscience Current Protocols in Neuroscience Pub Date : 2017-01-03 DOI:10.1002/cpns.21

Takako Kikkawa, Masanori Takahashi, Noriko Osumi

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引用次数: 1

摘要

本单元描述了使用胚胎期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，对体外培养的小鼠和大鼠胚胎的正常生长发育至关重要。还介绍了啮齿动物胚胎不同阶段的替代方案。由于WEC的胚胎是从子宫中解剖出来并在显微镜下操作的，因此可以克服使用子宫内电穿孔进行基因递送时遇到的许多困难。还包括以高度区域特异性的方式标记皮层原基的神经干/祖细胞的基因转移方法的描述。©2017 John Wiley&；股份有限公司。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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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.