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从人类肝脏切除标本中分离CD163库普弗细胞的方案。

Protocol for isolating CD163 Kupffer cells from human liver resections.

作者信息

Roca Suarez Armando Andres, Plissonnier Marie-Laure, Michelet Maud, Dubois Anaëlle, Heintz Sarah, Saez-Palma Maria, Delphin Marion, Bordes Isabelle, Molle Jennifer, Diederichs Audrey, Rodà Mélanie, Combe Emmanuel, Mougené Léa, Giraud Guillaume, Grand Xavier, Rivoire Michel, Levrero Massimo, Testoni Barbara, Zoulim Fabien

机构信息

INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France.

INSERM U1052, CNRS UMR-5286, Cancer Research Center of Lyon (CRCL), Lyon, France; University of Lyon, Université Claude-Bernard (UCBL), Lyon, France; The Lyon Hepatology Institute EVEREST, Lyon, France.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103359. doi: 10.1016/j.xpro.2024.103359. Epub 2024 Oct 4.

DOI:10.1016/j.xpro.2024.103359
PMID:39368096
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11491951/
Abstract

The liver microenvironment contains a wide variety of monocyte and macrophage populations. Here, we present a protocol for the specific isolation of liver-resident macrophages, known as Kupffer cells (KCs), from human liver resections. We describe steps for dissociating human liver tissues, separating non-parenchymal cells into fractions by a 2-phase iodixanol gradient, and positive selection of KCs based on the expression of CD163. We then provide instructions for validating the procedure by immunofluorescence to detect CD163. For complete details on the use and execution of this protocol, please refer to Roca Suarez, Plissonnier, et al..

摘要

肝脏微环境包含多种单核细胞和巨噬细胞群体。在此，我们展示了一种从人类肝脏切除组织中特异性分离肝脏驻留巨噬细胞（即库普弗细胞，KCs）的方案。我们描述了将人类肝脏组织解离、通过两相碘克沙醇梯度将非实质细胞分离成不同组分以及基于CD163表达对KCs进行阳性分选的步骤。然后，我们提供了通过免疫荧光检测CD163来验证该程序的说明。有关此方案的使用和执行的完整详细信息，请参考罗卡·苏亚雷斯、普利索尼耶等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/1463315c4631/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/f8b66ba69675/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/2cdfbc722402/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/4c138e6a787b/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/1463315c4631/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/f8b66ba69675/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/2cdfbc722402/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/4c138e6a787b/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76bf/11491951/1463315c4631/gr3.jpg

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本文引用的文献

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TLR8 agonist selgantolimod regulates Kupffer cell differentiation status and impairs HBV entry into hepatocytes via an IL-6-dependent mechanism.TLR8 激动剂 selgantolimod 通过一种依赖于 IL-6 的机制调节枯否细胞分化状态并损害 HBV 进入肝细胞。
Gut. 2024 Nov 11;73(12):2012-2022. doi: 10.1136/gutjnl-2023-331396.
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Single-cell, single-nucleus, and spatial transcriptomics characterization of the immunological landscape in the healthy and PSC human liver.单细胞、单细胞核和空间转录组学分析健康和 PSC 人肝中的免疫景观。
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Liver macrophages in health and disease.
肝脏巨噬细胞在健康和疾病中的作用。
Immunity. 2022 Sep 13;55(9):1515-1529. doi: 10.1016/j.immuni.2022.08.002.
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Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches.空间蛋白质组学揭示了独特且进化上保守的肝巨噬细胞生态位。
Cell. 2022 Jan 20;185(2):379-396.e38. doi: 10.1016/j.cell.2021.12.018. Epub 2022 Jan 11.
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Hepatitis B virus-induced modulation of liver macrophage function promotes hepatocyte infection.乙型肝炎病毒诱导的肝巨噬细胞功能调节促进肝细胞感染。
J Hepatol. 2019 Dec;71(6):1086-1098. doi: 10.1016/j.jhep.2019.06.032. Epub 2019 Jul 23.
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A human liver cell atlas reveals heterogeneity and epithelial progenitors.人类肝脏细胞图谱揭示了其异质性和上皮祖细胞。
Nature. 2019 Aug;572(7768):199-204. doi: 10.1038/s41586-019-1373-2. Epub 2019 Jul 10.
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Characterization of Pattern Recognition Receptor Expression and Functionality in Liver Primary Cells and Derived Cell Lines.鉴定肝脏原代细胞和衍生细胞系中模式识别受体的表达和功能。
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