Journal article

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

Johanna F Dekkers, Esmee 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, Hans Clevers, Anne C Rios



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 organs, such methods are lacking for breast tissue. Here we provide an optimized, highly versatile protocol for long-term culture of organoids derived from either normal human breast tissues or breast cancer (BC) tissues, as well as culturing conditions for a panel of 45 biobanked samples, including BC organoids covering all major disease subtypes (triple-negative, estrogen receptor-positive/progesterone receptor-positive and human epidermal g..

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University of Melbourne Researchers


Awarded by R35 grant from the National Cancer Institute

Awarded by European Research Council

Funding Acknowledgements

We are very grateful for the technical support of the Princess Maxima Center for Pediatric Oncology, the Hubrecht Institute, the Foundation Hubrecht Organoid Technology (HUB), the Dana-Farber Cancer Institute and the Walter and Eliza Hall Institute of Medical Research for generation of the described methods and the HUB for managing the breast organoid biobank. This work was financially supported by the Princess Maxima Center for Pediatric Oncology. J.F.D. was supported by a Marie Curie Global Fellowship and a VENI grant from the Dutch Organization for Scientific Research. J.M.R. and J.S.B. were supported by an R35 grant (CA242428) from the National Cancer Institute and acknowledge the support of the Susan G. Komen Breast Cancer Foundation. J.M.R. was supported by the American Cancer Society. J.E.V. was supported by the Australian National Health and Medical Research Council. A.C.R. was supported by a Starting Grant 2019 (project 804412) from the European Research Council.