Protocol for serial organoid formation assay using primary colorectal cancer tissues to evaluate cancer stem cell activity
Open accessAbstract
Organoids can enable the study of solid tumors initiated from a single cancer stem cell (CSC) ex vivo. We describe a serial tumor organoid plating protocol using primary colorectal cancer (CRC) tissues as a rapid and cost-efficient approach to evaluate the impact of therapeutic interventions on CSC functions. We detail the isolation of primary colorectal CSCs, organoid embedding, serial passaging, and CSC-related analytical techniques. For complete details on the use and execution of this protocol, please refer to Masibag et al. (2021) and Bergin et al. (2021).
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Tumor cells from biopsies or liquid samples can first be expanded in animal models as xenografts to obtain enough cells for organoid generation.
“Due to their low numbers, tumor cells from biopsies or liquid samples can be firstly expanded in animal models as xenografts in order to obtain sufficient cells for organoid generation57.”
From Organoids (2022) · cites this paper as [57]
The Step-by-step method details describe growing patient-derived cells as spheroids in culture before organoid formation, not expanding biopsy or liquid-sample cells as animal xenografts. The Limitations section mentions in vivo tumor transplantation only in connection with assessing therapeutic outcomes. Searches for xenograft expansion and biopsy or liquid samples found no passage supporting the attributed method; no page numbers were provided in our copy. (LLM, conf=0.76)
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- Authors
- Bergin, C. J., & Benoit, Y. D.
- Affiliations
- University of Ottawa (CA)
- Corresponding author
- Bergin, Christopher J.
- Benoit, Yannick D.
- Venue
- STAR Protocols
- Publisher
- Elsevier BV
- Published
- 2022-03
- Volume
- 3
- Issue
- 1
- Pages
- 101218
- ISSN
- 2666-1667 (ISSN-L)
- PMC
- PMC8899043
- OpenAlex
- W4214910533
- Cited by
- 14
- References
- 31
- Citations by year
- 2026: 3 · 2025: 3 · 2024: 5 · 2023: 2 · 2022: 1
- FWCI
- 1.15
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- STAR Protocols (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Ontario Ministry of Research and Innovation (ER17-13-012)
- Cancer Research Society (22778, 24039)
- Canadian Institutes of Health Research (PJT-173541)
- Natural Sciences and Engineering Research Council of Canada (DGECR-2018-00029, RGPIN-2018-06521)
- Ontario Ministry of Research, Innovation and Science
- Topics
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Keywords
- colorectal cancer, cancer stem cells, tumor organoids, primary tumor tissue
- MeSH terms
- Humans, Organoids, Neoplastic Stem Cells, Colorectal Neoplasms
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