Abstract:
The isolation and culturing of rodent retinal ganglion cells (RGC) is a key step in studying the function and cellular response of this crucial cell type. Typical methods used for isolation of RGCs include immunopanning or magnetic bead separation with antibodies targeting RGC specific protein markers. However, in developmental research, many of the most common markers, such as Thy-1, are not expressed in early stages of development. To help study these crucial early stage RGCs, we have developed a novel method that utilizes a transgenic mouse with a GFP tag on the protein BRN3 and a low-pressure fluorescence-activated cell sorter (FACS) system.
摘要:
啮齿动物视网膜神经节细胞(RGC)的分离和培养是研究这种关键细胞类型的功能和细胞反应的关键步骤。用于分离RGC的典型方法包括用靶向RGC特异性蛋白质标记的抗体进行免疫淘选或磁珠分离。然而,在发展研究中,许多最常见的标记,如Thy-1,在发展的早期阶段没有表达。为了帮助研究这些关键的早期RGC,我们开发了一种新方法,该方法利用在蛋白质BRN3上带有GFP标签的转基因小鼠和低压荧光激活细胞分选仪(FACS)系统。