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不使用基质胶从手术切除组织建立肺肿瘤类器官并进行表征的方案。

Protocol for the establishment and characterization of lung tumoroids from surgical resection without the use of Matrigel.

作者信息

Donà Federico, Erratico Silvia, Terracciano Luigi Maria, Brascia Debora, Cecconi Emanuela Re, Giudici Veronica Maria, Bossi Paola, Ng Charlotte K Y, Marulli Giuseppe, Piscuoglio Salvatore

机构信息

IRCCS Humanitas Research Hospital, Via Manzoni 56, Rozzano, 20089 Milan, Italy.

IRCCS Humanitas Research Hospital, Via Manzoni 56, Rozzano, 20089 Milan, Italy; Department of Biomedical Sciences, Humanitas University, Via Rita Levi Montalcini 4, Pieve Emanuele, 20072 Milan, Italy.

出版信息

STAR Protoc. 2025 Jul 2;6(3):103923. doi: 10.1016/j.xpro.2025.103923.

DOI:10.1016/j.xpro.2025.103923
PMID:40608510
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12269833/
Abstract

Organoids are vital for studying cancer mechanisms and treatment resistance, but their growth usually depends on costly, animal-derived Matrigel. Here, we present a protocol for expanding and maintaining lung-derived organoids without the need for Matrigel. We describe steps for establishing lung cancer organoids, staining organoids for immunohistochemistry, and cultivating them with ClinoStar. By eliminating dependence on Matrigel and its associated costs, this protocol enhances accessibility and scalability, making organoid-based research more feasible for cancer studies and therapeutic development.

摘要

类器官对于研究癌症机制和治疗抗性至关重要，但其生长通常依赖于昂贵的、动物来源的基质胶。在此，我们提出了一种无需基质胶即可扩增和维持肺来源类器官的方案。我们描述了建立肺癌类器官、对类器官进行免疫组织化学染色以及使用ClinoStar培养它们的步骤。通过消除对基质胶及其相关成本的依赖，该方案提高了可及性和可扩展性，使基于类器官的研究在癌症研究和治疗开发中更可行。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/22da3c5a6fd4/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/af897f7ff39a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/0165e68e3f0d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/8e6366cbb1de/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/f46bfcb507b6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/d9b3b777ab30/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/253f1e8e1677/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/22da3c5a6fd4/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/af897f7ff39a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/0165e68e3f0d/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/8e6366cbb1de/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/f46bfcb507b6/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/d9b3b777ab30/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/253f1e8e1677/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2ab1/12269833/22da3c5a6fd4/gr6.jpg

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本文引用的文献

1
Long-term expanding human airway organoids for disease modeling.长期扩增的人呼吸道类器官用于疾病建模。
EMBO J. 2019 Feb 15;38(4). doi: 10.15252/embj.2018100300. Epub 2019 Jan 14.
2
Comparison of Three Different TTF-1 Clones in Resected Primary Lung Cancer and Epithelial Pulmonary Metastases.切除原发性肺癌和肺上皮转移灶中三种不同 TTF-1 克隆的比较。
Am J Clin Pathol. 2018 Oct 24;150(6):533-544. doi: 10.1093/ajcp/aqy083.
3
p40 (ΔNp63) is superior to p63 for the diagnosis of pulmonary squamous cell carcinoma.p40（ΔNp63）在诊断肺鳞状细胞癌方面优于 p63。
Mod Pathol. 2012 Mar;25(3):405-15. doi: 10.1038/modpathol.2011.173. Epub 2011 Nov 4.