Organoid culture systems for prostate epithelial and cancer tissue

Abstract

Summary This protocol describes a recently developed strategy to generate 3D prostate organoid cultures from healthy mouse and human prostate (either bulk or FAC-sorted single luminal and basal cells), metastatic prostate cancer lesions and circulating tumour 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 tumour. The stepwise establishment of these cultures and the fully defined serum-free conditioned medium that is required to sustain organoid growth are outlined. Organoids established using this protocol can be used to study many different aspects of prostate biology, including homeostasis, tumorigenesis and drug discovery.

Targeting Bromodomain and Extra-Terminal (BET) Family Proteins in Castration-Resistant Prostate Cancer (CRPC)

“…Individual CP50 PDX experiments are detailed in Supplementary Methods. All animal studies were ethically reviewed and carried out in accordance with Animals (Scientific Procedures) Act 1986. previously described (42). PDOs were successfully generated from 22 of 36 (61%) metastatic CRPC biopsies between October 2015 and April 2016; nine of which were available for growth experiments.…”

A patient‐derived explant (PDE) model of hormone‐dependent cancer

“…The use of immortalized human cell line models for investigating novel therapies in vitro or in vivo is convenient but also a major reason for the high failure of new drugs entering clinical trials. To move toward more clinically relevant model systems, researchers have adopted patient‐derived approaches such as organoids (Drost et al ., 2016; Gao et al ., 2014) and xenografts (PDX) (Lawrence et al ., 2013; Wang et al ., 2005; Whittle et al ., 2015). We report an alternative approach of culturing freshly resected breast and prostate cancer tissue as patient‐derived explants (PDE).…”

Towards personalized computational oncology: from spatial models of tumour spheroids, to organoids, to tissues

“…These cultures serve as an experimental model of normal organ development. Various non-tumorigenic cell lines have been grown as hollow acini, including those originating from the breast [12], kidney [13], lung [14], pancreas [15] or prostate [16]. Such 3D cultures have also been used to examine changes in both the growth and morphology of non-tumorigenic cells in altered microenvironments [17,18], to delineate the oncogenetic mechanisms of mutated cells [19,20] and to test the response of tumour cells to various treatments [21,22].…”