分析利用血脑屏障体外模型中转录因子布鲁斯菌迁移效率的方案。
Protocol to analyze the transmigration efficiency of T. brucei using an in vitro model of the blood-cerebrospinal fluid barrier.
机构信息
Interfaculty Institute of Biochemistry, University of Tübingen, Tübingen, Germany.
Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba, Tsukuba, Japan.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101676. doi: 10.1016/j.xpro.2022.101676. Epub 2022 Sep 13.
At present, the only approach to investigate the transmigration of Trypanosoma brucei, the causative agent of human African trypanosomiasis, from blood to cerebrospinal fluid is through animal experiments. This protocol details how to analyze the transmigration efficiency using an in vitro model of the blood-cerebrospinal fluid (blood-CSF) barrier. We describe how to grow human choroid plexus epithelial cells on cell culture filter inserts to form the barrier, followed by isolating and quantifying genomic DNA of transmigrated parasites by qPCR. For complete details on the use and execution of this protocol, please refer to Speidel et al. (2022).
目前，研究引起人类非洲锥虫病的布氏锥虫从血液到脑脊液迁移的唯一方法是通过动物实验。本方案详细介绍了如何使用体外血脑屏障（blood-CSF）模型分析迁移效率。我们描述了如何在细胞培养过滤器插入物上培养人脉络丛上皮细胞以形成屏障，然后通过 qPCR 分离和定量迁移寄生虫的基因组 DNA。有关该方案使用和执行的完整详细信息，请参阅 Speidel 等人（2022 年）。
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