Levy Rachelle, Molina Jessica, Danielpour Moise, Breunig Joshua J
Department of Neurosurgery, Regenerative Medicine Institute, Cedars-Sinai Medical Center.
Department of Biomedical Sciences, Regenerative Medicine Institute, Cedars-Sinai Medical Center.
J Vis Exp. 2014 May 7(87):51319. doi: 10.3791/51319.
Over the past several years the pial surface has been identified as a germinal niche of importance during embryonic, perinatal and adult neuro- and gliogenesis, including after injury. However, methods for genetically interrogating these progenitor populations and tracking their lineages had been limited owing to a lack of specificity or time consuming production of viruses. Thus, progress in this region has been relatively slow with only a handful of investigations of this location. Electroporation has been used for over a decade to study neural stem cell properties in the embryo, and more recently in the postnatal brain. Here we describe an efficient, rapid, and simple technique for the genetic manipulation of pial surface progenitors based on an adapted electroporation approach. Pial surface electroporation allows for facile genetic labeling and manipulation of these progenitors, thus representing a time-saving and economical approach for studying these cells.
在过去的几年里，软脑膜表面已被确定为在胚胎期、围产期和成年期神经及神经胶质生成过程中（包括损伤后）重要的生发微环境。然而，由于缺乏特异性或病毒生产耗时，对这些祖细胞群体进行基因询问和追踪其谱系的方法一直有限。因此，该领域的进展相对缓慢，对此部位的研究仅有少数几项。电穿孔已被用于研究胚胎期以及最近产后大脑中的神经干细胞特性超过十年。在此，我们描述了一种基于改良电穿孔方法对软脑膜表面祖细胞进行基因操作的高效、快速且简单的技术。软脑膜表面电穿孔能够方便地对这些祖细胞进行基因标记和操作，因此是一种研究这些细胞的省时且经济的方法。
