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Nat Protoc. 2016 Feb;11(2):347-58. doi: 10.1038/nprot.2016.006. Epub 2016 Jan 21.

Organoid culture systems for prostate epithelial and cancer tissue.

Author information

1
Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht, Utrecht, the Netherlands.
2
Cancer Genomics Netherlands, UMC Utrecht, Utrecht, the Netherlands.
3
Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, New York, USA.
4
Department of Medicine, Weill Cornell Medical College and New York-Presbyterian Hospital, New York, New York, USA.
5
Department of Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA.

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.

PMID:
26797458
PMCID:
PMC4793718
DOI:
10.1038/nprot.2016.006
[Indexed for MEDLINE]
Free PMC Article
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