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生成巨噬细胞与乳腺癌类肿瘤共培养物的方案。

Protocol for generating a co-culture of macrophages with breast cancer tumoroids.

作者信息

Raffo-Romero Antonella, Ziane-Chaouche Lydia, Hajjaji Nawale, Salzet Michel, Duhamel Marie

机构信息

University Lille, Inserm, CHU Lille, U1192 - Protéomique Réponse Inflammatoire Spectrométrie de Masse - PRISM, F-59000 Lille, France; Equipe Labellisée Ligue Contre le Cancer, Lille, France.

University Lille, Inserm, CHU Lille, U1192 - Protéomique Réponse Inflammatoire Spectrométrie de Masse - PRISM, F-59000 Lille, France; Equipe Labellisée Ligue Contre le Cancer, Lille, France.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103536. doi: 10.1016/j.xpro.2024.103536. Epub 2024 Dec 20.

DOI:10.1016/j.xpro.2024.103536
PMID:39709607
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11726783/
Abstract

Cancer progression and treatment outcomes are heavily influenced by the tumor microenvironment (TME), especially through immune cell interactions. Here, we present a protocol for generating co-cultures of tumoroids with macrophages, either semi-liquid or Matrigel-embedded. We describe steps for macrophage preparation, co-culture establishment, and medium adjustments to support cell viability and function. While optimized for breast cancer models, this protocol can be adapted to other tumor types with appropriate medium adjustments. For complete details on the use and execution of this protocol, please refer to Raffo-Romero et al..

摘要

癌症进展和治疗结果受到肿瘤微环境(TME)的严重影响,尤其是通过免疫细胞相互作用。在此,我们展示了一种用于生成肿瘤类器官与巨噬细胞共培养物的方案,共培养物可以是半液体形式或包埋于基质胶中。我们描述了巨噬细胞制备、共培养建立以及培养基调整的步骤,以支持细胞活力和功能。虽然该方案是针对乳腺癌模型进行优化的,但通过适当调整培养基,也可适用于其他肿瘤类型。有关该方案使用和执行的完整详细信息,请参考拉福 - 罗梅罗等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/5bf9bdcb6b48/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/53f85e8b9f4a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/e2ddb3908394/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/dcfa1b0c4ce7/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/6801e1e7e76c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/3c4869109510/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/3f71c4f45222/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/5bf9bdcb6b48/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/53f85e8b9f4a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/e2ddb3908394/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/dcfa1b0c4ce7/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/6801e1e7e76c/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/3c4869109510/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/3f71c4f45222/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d207/11726783/5bf9bdcb6b48/gr6.jpg

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本文引用的文献

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Cell Rep Methods. 2024 Jun 17;4(6):100792. doi: 10.1016/j.crmeth.2024.100792. Epub 2024 Jun 10.
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Protocol for generation of and high-throughput drug testing with patient-derived colorectal cancer organoids.患者来源结直肠癌细胞类器官的生成及高通量药物筛选的方案。
STAR Protoc. 2024 Jun 21;5(2):103090. doi: 10.1016/j.xpro.2024.103090. Epub 2024 May 28.
3
Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids.
人正常和乳腺癌类器官的长期培养、遗传操作和异种移植。
Nat Protoc. 2021 Apr;16(4):1936-1965. doi: 10.1038/s41596-020-00474-1. Epub 2021 Mar 10.
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Paclitaxel Treatment and Proprotein Convertase 1/3 (PC1/3) Knockdown in Macrophages is a Promising Antiglioma Strategy as Revealed by Proteomics and Cytotoxicity Studies.紫杉醇治疗和巨噬细胞中前蛋白转化酶 1/3(PC1/3)敲低通过蛋白质组学和细胞毒性研究揭示了一种有前途的抗神经胶质瘤策略。
Mol Cell Proteomics. 2018 Jun;17(6):1126-1143. doi: 10.1074/mcp.RA117.000443. Epub 2018 Mar 12.
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The prognostic landscape of genes and infiltrating immune cells across human cancers.人类癌症中基因与浸润性免疫细胞的预后情况
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