Kikkawa Takako, Takahashi Masanori, Osumi Noriko
Department of Developmental Neuroscience, United Center for Advanced Research and Translational Medicine (ART), Tohoku University School of Medicine, Sendai, Miyagi, Japan.
Jichi Medical University Graduate School of Medicine, Shimotsuke, Tochigi, Japan.
Curr Protoc Neurosci. 2017 Jan 3;78:3.30.1-3.30.16. doi: 10.1002/cpns.21.
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.
本单元描述了使用胚胎第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，包括制备常用于WEC的高质量即时离心（IC）大鼠血清，该血清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。还介绍了针对不同阶段啮齿动物胚胎的替代方案。由于用于WEC的胚胎是从子宫中取出并在显微镜下操作的，因此可以克服使用子宫内电穿孔时遇到的许多基因传递困难。还包括一种以高度区域特异性方式标记皮质原基神经干细胞/祖细胞的基因转移方法的描述。©2017约翰威立父子公司。
