Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids

Hans Clevers, Johanna F. Dekkers, Esmée J. van Vliet, Norman Sachs, Jennifer M. Rosenbluth, Oded Kopper, Heggert G. Rebel, Ellen J. Wehrens, Carol Piani, Jane E. Visvader, Carla S. Verissimo, Sylvia F. Boj, Joan S. Brugge, Anne C. Rios

Published: 2021-03-09 DOI: 10.1038/s41596-020-00474-1

xenotransplantation

breast cancer organoids

long-term culture

genetic manipulation

orthotopic transplantation

AI 解读

Generation of mouse–human chimeric embryos

Genome-wide quantification of transcription factor binding at single-DNA-molecule resolution using methyl-transferase footprinting

Generation of CRISPR–Cas9-mediated genetic knockout human intestinal tissue–derived enteroid lines by lentivirus transduction and single-cell cloning

A puromycin-dependent activity-based sensing probe for histochemical staining of hydrogen peroxide in cells and animal tissues

Decoding personal biotic and abiotic airborne exposome

A clinically relevant model of focal embolic cerebral ischemia by thrombus and thrombolysis in rhesus monkeys

Bioengineering human intestinal mucosal grafts using patient-derived organoids, fibroblasts and scaffolds

Biomimetic tail-to-head terpene cyclizations using the resorcin[4]arene capsule catalyst

Neuroscience robotics for controlled induction and real-time assessment of hallucinations

Scanorama: integrating large and diverse single-cell transcriptomic datasets

Single-cell RNA sequencing (scRNA-seq)（单细胞RNA测序）

Data integration（数据集成）

Heterogeneity（异质性）

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Organoid technology has revolutionized the study of human organ development, disease and therapy response tailored to the individual. Although detailed protocols are available for the generation and long-term propagation of human organoids from various or
Introduction
Materials
Procedure
Troubleshooting
Timing
Anticipated results
- Reporting Summary
- References
References
ACKNOWLEDGMENTS

DeSantis, C. E. et al. Breast cancer statistics, 2019. CA Cancer J. Clin. 69, 438–451 (2019).
Nik-Zainal, S. et al. Landscape of somatic mutations in 560 breast cancer whole-genome sequences. Nature 534, 47–54 (2016).
Curtis, C. et al. The genomic and transcriptomic architecture of 2,000 breast tumours reveals novel subgroups. Nature 486, 346–352 (2012).
Rios, A. C. et al. Intraclonal plasticity in mammary tumors revealed through large-scale single-cell resolution 3D imaging. Cancer Cell 35, 618–632 (2019).
Hoon Tan, P. et al The 2019 World Health Organization classification of tumours of the breast. Histopathology 77, 181–185 (2020).
Sato, T. et al. Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459, 262–265 (2009).
Sachs, N. et al. A living biobank of breast cancer organoids captures disease heterogeneity. Cell 172, 373–386 (2018).
Rosenbluth, J. M. et al. Organoid cultures from normal and cancer-prone human breast tissues preserve complex epithelial lineages. Nat. Commun. 11, 1711 (2020).
Dekkers, J. F. et al. Modeling breast cancer using CRISPR–Cas9-mediated engineering of human breast organoids. J. Natl. Cancer Inst 112, 540–544 (2020).
Kedrin, D. et al. Intravital imaging of metastatic behavior through a mammary imaging window. Nat. Methods 5, 1019–1021 (2008).
Holliday, D. L. & Speirs, V. Choosing the right cell line for breast cancer research. Breast Cancer Res. 13, 76–99 (2011).
Kapałczyńska, M. et al. 2D and 3D cell cultures—a comparison of different types of cancer cell cultures. Arch. Med. Sci. 14, 910–919 (2018).
Vargo-Gogola, T. & Rosen, J. M. Modelling breast cancer: one size does not fit all. Nat. Rev. Cancer 7, 659–672 (2007).
Neve, R. M. et al. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell 10, 515–527 (2006).
Miller, D. H., Sokol, E. S. & Gupta, P. B. 3D primary culture model to study human mammary development. Methods Mol. Biol. 1612, 139–147 (2017).
Linnemann, J. R., Meixner, L. K., Miura, H. & Scheel, C. H. An organotypic 3D assay for primary human mammary epithelial cells that recapitulates branching morphogenesis. Methods Mol. Biol. 1612, 125–137 (2017).
Lim, E. et al. Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers. Nat. Med. 15, 907–913 (2009).
Kopper, O. et al. An organoid platform for ovarian cancer captures intra- and interpatient heterogeneity. Nat. Med. 25, 838–849 (2019).
Cattaneo, C. M. et al. Tumor organoid–T-cell coculture systems. Nat. Protoc. 15, 15–39 (2020).
Drost, J. et al. Organoid culture systems for prostate epithelial and cancer tissue. Nat. Protoc. 11, 347–358 (2016).
Fujii, M., Matano, M., Nanki, K. & Sato, T. Efficient genetic engineering of human intestinal organoids using electroporation. Nat. Protoc. 10, 1474–1485 (2015).
Broutier, L. et al. Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat. Protoc. 11, 1724–1743 (2016).
Drost, J. et al. Sequential cancer mutations in cultured human intestinal stem cells. Nature 521, 43–47 (2015).
Dall, G., Vieusseux, J., Unsworth, A., Anderson, R. & Britt, K. Low dose, low cost estradiol pellets can support MCF-7 tumour growth in nude mice without bladder symptoms. J. Cancer 6, 1331–1336 (2015).
Dobrolecki, L. E. et al. Patient-derived xenograft (PDX) models in basic and translational breast cancer research. Cancer Metastasis Rev. 35, 547–573 (2016).
Gakhar, G., Wight-Carter, M., Andrews, G., Olson, S. & Nguyen, T. A. Hydronephrosis and urine retention in estrogen-implanted athymic nude mice. Vet. Pathol. 46, 505–508 (2009).
Whittle, J. R. et al. Dual targeting of CDK4/6 and BCL2 pathways augments tumor response in estrogen receptor positive breast cancer. Clin. Cancer Res. 26, 4120–4134 (2020).

Long-term culture, genetic manipulation and xenotransplantation of human normal and breast cancer organoids

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