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Organoid culture systems for prostate epithelial and cancer tissue

Nature Protocols volume 11, pages 347–358 (2016)Cite this article

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Abstract

This protocol describes a strategy for the generation of 3D prostate organoid cultures from healthy mouse and human prostate cells (either bulk or FACS-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumor cells. Organoids derived from healthy material contain the differentiated luminal and basal cell types, whereas organoids derived from prostate cancer tissue mimic the histology of the tumor. We explain how to establish these cultures in the fully defined serum-free conditioned medium that is required to sustain organoid growth. Starting with the plating of digested tissue material, full-grown organoids can usually be obtained in ∼2 weeks. The culture protocol we describe here is currently the only one that allows the growth of both the luminal and basal prostatic epithelial lineages, as well as the growth of advanced prostate cancers. Organoids established using this protocol can be used to study many different aspects of prostate biology, including homeostasis, tumorigenesis and drug discovery.

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Figure 1: Dissection of the mouse prostate.
Figure 2: Establishment of mouse prostate organoid cultures.
Figure 3: Establishment of mouse and human luminal cell– and basal cell–derived prostate organoid cultures.

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Acknowledgements

We thank members of the contributing laboratories for support. We are grateful for support from the following: The Netherlands Organisation for Scientific Research (NWO-ZonMw) VENI grant to J.D. (91614138); Koningin Wilhelmina Fonds (Dutch cancer foundation) to W.R.K. (2015-7545); the Prostate Cancer Foundation (C.L.S., Y.C.); and the CancerGenomics.nl (NWO Gravitation) program.

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Author notes

  1. Jarno Drost and Wouter R Karthaus: These authors contributed equally to this work.

Authors and Affiliations

  1. Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht, Utrecht, the Netherlands

    Jarno Drost, Else Driehuis & Hans Clevers

  2. Cancer Genomics Netherlands, UMC Utrecht, Utrecht, the Netherlands

    Jarno Drost, Else Driehuis & Hans Clevers

  3. Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Wouter R Karthaus, Dong Gao, Charles L Sawyers & Yu Chen

  4. Department of Medicine, Weill Cornell Medical College and New York-Presbyterian Hospital, New York, New York, USA

    Yu Chen

  5. Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Yu Chen

Authors
  1. Jarno Drost
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  2. Wouter R Karthaus
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  3. Dong Gao
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  4. Else Driehuis
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  5. Charles L Sawyers
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  6. Yu Chen
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  7. Hans Clevers
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Contributions

J.D., W.R.K., Y.C., C.L.S. and H.C. conceived the study. J.D., W.R.K., D.G. and E.D. wrote the manuscript.

Corresponding authors

Correspondence to Jarno Drost or Hans Clevers.

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The authors declare no competing financial interests.

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Drost, J., Karthaus, W., Gao, D. et al. Organoid culture systems for prostate epithelial and cancer tissue. Nat Protoc 11, 347–358 (2016). https://doi.org/10.1038/nprot.2016.006

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