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Organoid culture systems for prostate epithelial and cancer tissue


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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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 (NWO Gravitation) program.

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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.

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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).

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