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体外细胞培养模型中从吞噬耐甲氧西林金黄色葡萄球菌的肺泡巨噬细胞分离细胞外囊泡的方案。

Protocol for isolating extracellular vesicles from alveolar macrophages phagocytosing MRSA in vitro cell culture models.

作者信息

Bai Songjie, Hu Ruomeng, Wen Xuehuan, Zhang Wenting, Li Bingyu, Yang Jie, Yu Qing, Zeng Xianchang, Xiu Huiqing, Cai Zhijian, Zhang Gensheng

机构信息

Department of Critical Care Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, China; Department of Cardiovascular Surgery, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, China.

Department of Critical Care Medicine, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang 310009, China.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103526. doi: 10.1016/j.xpro.2024.103526. Epub 2024 Dec 20.

DOI:10.1016/j.xpro.2024.103526
PMID:39708327
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11730243/
Abstract

Extracellular vesicles (EVs) play a crucial role in delivering bioactive cargo in infectious diseases. Here, we present a protocol for isolating EVs from alveolar macrophages (AMs) that phagocytose methicillin-resistant Staphylococcus aureus (MRSA) in vitro cell culture models. We describe steps for bacterial preparation; infection of AMs with MRSA; and isolation, purification, and characterization of EVs. This protocol provides a valuable perspective for studying EVs derived from pathogen-infected immune cells. For complete details on the use and execution of this protocol, please refer to Bai et al..

摘要

细胞外囊泡(EVs)在传染病中生物活性物质的传递过程中起着关键作用。在此,我们介绍一种从体外细胞培养模型中吞噬耐甲氧西林金黄色葡萄球菌(MRSA)的肺泡巨噬细胞(AMs)中分离EVs的方法。我们描述了细菌制备、AMs用MRSA感染以及EVs的分离、纯化和表征的步骤。该方法为研究源自病原体感染免疫细胞的EVs提供了有价值的视角。有关该方法的使用和实施的完整详细信息,请参考Bai等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/056fc95df944/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/2ce330c8964d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/90e09aa9c3c1/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/db7e02da188f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/f75399f30bd7/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/056fc95df944/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/2ce330c8964d/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/90e09aa9c3c1/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/db7e02da188f/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/f75399f30bd7/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ac8/11730243/056fc95df944/gr4.jpg

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Protocol for isolating extracellular vesicles from alveolar macrophages phagocytosing MRSA in vitro cell culture models.体外细胞培养模型中从吞噬耐甲氧西林金黄色葡萄球菌的肺泡巨噬细胞分离细胞外囊泡的方案。
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本文引用的文献

1
Extracellular vesicles from alveolar macrophages harboring phagocytosed methicillin-resistant Staphylococcus aureus induce necroptosis.肺泡巨噬细胞来源的含有吞噬的耐甲氧西林金黄色葡萄球菌的细胞外囊泡诱导细胞坏死。
Cell Rep. 2024 Jul 23;43(7):114453. doi: 10.1016/j.celrep.2024.114453. Epub 2024 Jul 9.
2
Extracellular vesicles derived from M2-like macrophages alleviate acute lung injury in a miR-709-mediated manner.M2 样巨噬细胞来源的细胞外囊泡通过 miR-709 介导的方式缓解急性肺损伤。
J Extracell Vesicles. 2024 Apr;13(4):e12437. doi: 10.1002/jev2.12437.
3
Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches.
细胞外囊泡研究的最低信息要求(MISEV2023):从基础到先进方法。
J Extracell Vesicles. 2024 Feb;13(2):e12404. doi: 10.1002/jev2.12404.
4
Exosomal lipid PI4P regulates small extracellular vesicle secretion by modulating intraluminal vesicle formation.外泌体脂质 PI4P 通过调节腔内囊泡形成来调节小细胞外囊泡的分泌。
J Extracell Vesicles. 2023 Apr;12(4):e12319. doi: 10.1002/jev2.12319.
5
Comprehensive isolation of extracellular vesicles and nanoparticles.细胞外囊泡和纳米颗粒的综合分离。
Nat Protoc. 2023 May;18(5):1462-1487. doi: 10.1038/s41596-023-00811-0. Epub 2023 Mar 13.
6
Macrophage-derived exosomal aminopeptidase N aggravates sepsis-induced acute lung injury by regulating necroptosis of lung epithelial cell.巨噬细胞衍生的外泌体氨基肽酶 N 通过调节肺上皮细胞坏死性凋亡加重脓毒症诱导的急性肺损伤。
Commun Biol. 2022 Jun 6;5(1):543. doi: 10.1038/s42003-022-03481-y.
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LPS-induced macrophage HMGB1-loaded extracellular vesicles trigger hepatocyte pyroptosis by activating the NLRP3 inflammasome.脂多糖诱导的巨噬细胞高迁移率族蛋白B1负载的细胞外囊泡通过激活NLRP3炎性小体触发肝细胞焦亡。
Cell Death Discov. 2021 Nov 6;7(1):337. doi: 10.1038/s41420-021-00729-0.
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The clinical role of host and bacterial-derived extracellular vesicles in pneumonia.宿主和细菌来源的细胞外囊泡在肺炎中的临床作用。
Adv Drug Deliv Rev. 2021 Sep;176:113811. doi: 10.1016/j.addr.2021.05.021. Epub 2021 May 20.
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Extracellular vesicles: An emerging platform in gram-positive bacteria.细胞外囊泡:革兰氏阳性菌中的一个新兴平台。
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