体外诱导人平滑肌细胞和内皮细胞固有免疫记忆的方案。
Protocol for the induction of innate immune memory in human smooth muscle cells and endothelial cells in vitro.
机构信息
Department of Cardiology I - Coronary and Peripheral Vascular Disease, Heart Failure, University Hospital Münster, Münster, Germany.
Department of Cardiology I - Coronary and Peripheral Vascular Disease, Heart Failure, University Hospital Münster, Münster, Germany; Department of Medical Genetics, Third Faculty of Medicine, Charles University, Prague, Czechia; Institutes of Molecular Genetics of the Czech Academy of Sciences, Prague, Czechia.
出版信息
STAR Protoc. 2024 Mar 15;5(1):102776. doi: 10.1016/j.xpro.2023.102776. Epub 2023 Dec 19.
Non-immune cells, like innate immune cells, can develop a memory-like phenotype in response to priming with microbial compounds or certain metabolites, which enables an enhanced response to a secondary unspecific stimulus. This paper describes a step-by-step protocol for the induction and analysis of trained immunity in human endothelial and smooth muscle cells. We then describe steps for cell culture with cryopreserved vascular cells, subcultivation, and induction of trained immunity. We then provide detailed procedures for downstream analysis using ELISA and qPCR. For complete details on the use and execution of this protocol, please refer to Sohrabi et al. (2020) and Shcnack et al..
非免疫细胞，如先天免疫细胞，在受到微生物化合物或某些代谢物的启动后，可以发展出类似记忆的表型，从而增强对二次非特异性刺激的反应。本文描述了在人内皮和平滑肌细胞中诱导和分析训练免疫的逐步方案。然后，我们描述了使用冷冻保存的血管细胞进行细胞培养、传代和诱导训练免疫的步骤。然后，我们提供了使用 ELISA 和 qPCR 进行下游分析的详细程序。有关此方案的使用和执行的完整详细信息，请参阅 Sohrabi 等人（2020 年）和 Shcnack 等人。
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