Elias Laura, Kriegstein Arnold
Institute for Regeneration Medicine, University of California, San Francisco, USA.
J Vis Exp. 2007(6):235. doi: 10.3791/235. Epub 2007 Jul 11.
Organotypic slice cultures from embryonic rodent brains are widely used to study brain development. While there are often advantages to an in-vivo system, organotypic slice cultures allow one to perform a number of manipulations that are not presently feasible in-vivo. To date, organtotypic embryonic brain slice cultures have been used to follow individual cells using time-lapse microscopy, manipulate the expression of genes in the ganglionic emanances (a region that is hard to target by in-utero electroporation), as well as for pharmacological studies. In this video protocol we demonstrate how to make organotypic slice cultures from rat embryonic day 18 embryos. The protocol involves dissecting the embryos, embedding them on ice in low melt agarose, slicing the embedded brains on the vibratome, and finally plating the slices onto filters in culture dishes. This protocol is also applicable in its present form to making organotypic slice cultures from different embryonic ages for both rats and mice.
来自胚胎期啮齿动物大脑的器官型切片培养物被广泛用于研究大脑发育。虽然体内系统通常有其优势，但器官型切片培养允许人们进行一些目前在体内无法实现的操作。迄今为止，器官型胚胎脑切片培养物已被用于通过延时显微镜追踪单个细胞、操纵神经节发出部位（一个难以通过子宫内电穿孔靶向的区域）的基因表达，以及用于药理学研究。在本视频方案中，我们展示了如何从大鼠胚胎第18天的胚胎制作器官型切片培养物。该方案包括解剖胚胎、将它们在冰上嵌入低熔点琼脂糖、在振动切片机上切割嵌入的大脑，最后将切片接种到培养皿中的滤膜上。该方案以其目前的形式也适用于从不同胚胎年龄的大鼠和小鼠制作器官型切片培养物。
