Establishment of patient-derived cancer organoids for drug-screening applications

被引：480

作者：

Driehuis, Else ^{[1
,2
,3
,4
]}

Kretzschmar, Kai ^{[1
,2
,5
]}

Clevers, Hans ^{[1
,2
,6
]}

机构：

[1] Royal Netherlands Acad Arts & Sci KNAW, Hubrecht Inst, Oncode Inst, Utrecht, Netherlands

[2] Univ Med Ctr UMC Utrecht, Utrecht, Netherlands

[3] Hubrecht Organoid Technol HUB, Utrecht, Netherlands

[4] Univ Med Ctr UMC Utrecht, Dept Pathol, Utrecht, Netherlands

[5] Univ Hosp Wurzburg, Mildred Scheel Early Career Ctr MSNZ Canc Res, Wurzburg, Germany

[6] Princess Maxima Ctr Pediat Oncol, Utrecht, Netherlands

来源：

NATURE PROTOCOLS | 2020年 / 15卷 / 10期

基金：

欧洲研究理事会;

关键词：

IN-VITRO EXPANSION; STEM-CELL NICHE; COLORECTAL-CANCER; SINGLE LGR5; DISEASE; MODELS; INHIBITION; CULTURE; MOUSE; LIVER;

D O I：

10.1038/s41596-020-0379-4

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Adult stem cell-based organoid technology is a versatile tool for the generation and long-term maintenance of near-native 3D epithelial tissues in vitro. The generation of cancer organoids from primary patient material enables a range of therapeutic agents to be tested in the resulting organoid cultures. Patient-derived cancer organoids therefore hold great promise for personalized medicine. Here, we provide an overview of the protocols used by different groups to establish organoids from various epithelial tissues and cancers, plus the different protocols subsequently used to test the in vitro therapy sensitivity of these patient-derived organoids. We also provide an in-depth protocol for the generation of head and neck squamous cell carcinoma organoids and their subsequent use in semi-automated therapy screens. Establishment of organoids and subsequent screening can be performed within 3 months, although this timeline is highly dependent on a.o. starting material and the number of therapies tested. The protocol provided may serve as a reference to successfully establish organoids from other cancer types and perform drug screenings thereof. This protocol summarizes the various approaches available to derive organoids from cancer patients and use these for screening of possible treatments. An optimized protocol for using head and neck cancer organoids is also described.

引用

页码：3380 / 3409

页数：30

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