Organoid culture systems for prostate epithelial and cancer tissue

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.

Figure 1. Establishment of mouse prostate organoid cultures

A) Overview of the isolation of the prostate from the mouse urogenital system (for a detailed isolation protocol see). The procedure refers to pictures I – IX. B) Schematic representation of the anatomy of the mouse prostate. AP, anterior prostate; DLP, dorsolateral prostate; VP, ventral prostate. C) Example of how to plate the matrigel disc in a well of a tissue culture plate. D) Representative pictures of organoids growing from mouse prostate tissue 1 and 7 days after plating. Scale bars, 100 micron.

Figure 2. Establishment of mouse and human luminal and basal cell-derived prostate organoid cultures

A) Overview of the establishment of luminal and basal-cell derived mouse prostate organoid cultures using FACS. Cells are sorted based on expression of Cd24 (luminal cells) and Cd49f (basal cells). Scale bars, 100 micron. B) Overview of the establishment of luminal and basal-cell derived human prostate organoid cultures using FACS. Cells are sorted based on expression of CD26 (luminal cells) and CD49f (basal cells). Scale bars, 100 micron.