Abstract：
Mounting evidence indicates that the immune response triggered by brain disorders (e.g., brain ischemia and autoimmune encephalomyelitis) occurs not only in the brain, but also in the skull. A key step toward analyzing changes in immune cell populations in both the brain and skull bone marrow after brain damage (e.g., stroke) is to obtain sufficient numbers of high-quality immune cells for downstream analyses. Here, two optimized protocols are provided for isolating immune cells from the brain and skull bone marrow. The advantages of both protocols are reflected in their simplicity, speed, and efficacy in yielding a large quantity of viable immune cells. These cells may be suitable for a range of downstream applications, such as cell sorting, flow cytometry, and transcriptomic analysis. To demonstrate the effectiveness of the protocols, immunophenotyping experiments were performed on stroke brains and normal brain skull bone marrow using flow cytometry analysis, and the results aligned with findings from published studies.
摘要：
越来越多的证据表明，由脑部疾病引发的免疫反应(例如，脑缺血和自身免疫性脑脊髓炎）不仅发生在大脑中，还有头骨.分析脑损伤后大脑和颅骨骨髓中免疫细胞群变化的关键步骤（例如，中风)是为了获得足够数量的高质量免疫细胞用于下游分析。这里,提供了两种优化的方案，用于从脑和颅骨骨髓中分离免疫细胞。两种协议的优点都体现在其简单性，速度,以及产生大量活免疫细胞的功效。这些细胞可能适用于一系列下游应用，比如细胞分选，流式细胞术,和转录组学分析。为了证明协议的有效性，使用流式细胞术分析对中风脑和正常脑颅骨骨髓进行免疫表型分析实验，结果与已发表的研究结果一致。
