Miglietta Audrey G, David-Bercholz Jennifer, Munji Roeben N, Pulido Robert S, Soung Allison L, Terrando Niccolò, Daneman Richard, Yang Ting
Departments of Pharmacology and Neurosciences, University of California, San Diego, La Jolla, CA 92093, USA; Neurosciences Graduate Program, University of California, San Diego, La Jolla, CA 92093, USA.
Department of Anesthesiology, Center for Translational Pain Medicine, Duke University, Durham, NC 27710, USA.
STAR Protoc. 2025 Jun 20;6(2):103837. doi: 10.1016/j.xpro.2025.103837. Epub 2025 May 21.
Brain endothelial cells (ECs) line blood vessel walls and form the blood-brain barrier (BBB), facilitating interactions between the brain and the periphery. Here, we present a protocol for isolating purified ECs from adult mouse brains using fluorescence-activated cell sorting (FACS) for downstream analyses, such as RNA sequencing. We describe the steps for brain dissection, enzymatic tissue digestion, trituration, myelin removal, and FACS. Furthermore, we detail adaptations of this protocol for isolating ECs from brain subregions, such as the hippocampus. For complete details on the use and execution of this protocol, please refer to Munji et al. and Pulido et al..
脑内皮细胞（ECs）排列在血管壁上，形成血脑屏障（BBB），促进大脑与外周之间的相互作用。在这里，我们展示了一种使用荧光激活细胞分选（FACS）从成年小鼠大脑中分离纯化ECs的方案，用于下游分析，如RNA测序。我们描述了脑解剖、酶促组织消化、研磨、髓磷脂去除和FACS的步骤。此外，我们详细说明了该方案在从脑亚区域（如海马体）分离ECs方面的调整。有关该方案使用和执行的完整详细信息，请参考Munji等人和Pulido等人的研究。
