Single luminal epithelial progenitors can generate prostate organoids in culture

Abstract

The intrinsic ability to display self-organizing morphogenetic properties in ex vivo culture may represent a general property of tissue stem cells. Here we show that single luminal stem/progenitor cells can generate prostate organoids in a three-dimensional culture system in the absence of stroma. Organoids generated from CARNs (castration-resistant Nkx3.1-expressing cells) or normal prostate epithelium exhibit tissue architecture containing luminal and basal cells, undergo long-term expansion in culture, and display functional androgen receptor signaling. Lineage-tracing demonstrates that luminal cells are favored for organoid formation, and generate basal cells in culture. Furthermore, tumor organoids can initiate from CARNs after oncogenic transformation, and from mouse models of prostate cancer, and can facilitate analyses of drug response. Finally, we provide evidence supporting the feasibility of organoid studies of human prostate tissue. Our studies underscore the progenitor properties of luminal cells, and identify in vitro approaches for studying prostate biology.

Prostate epithelial cell of origin determines cancer differentiation state in an organoid transformation assay

“…Hi basal cell 1,000 21 (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29)(30) Intriguingly, human basal-and luminal-derived tumors demonstrate histologically distinct phenotypes when challenged with the same oncogenes, MYC and AKT1. Tumors derived from basal cells are of a higher grade than tumors derived from luminal cells.…”

Prostate Sphere-forming Stem Cells Are Derived from the P63-expressing Basal Compartment

“…Although it has been reported that prostaspheres expressed basal cell cytokeratins and P63, the precise placement of prostasphere cells in the overall prostate stem cell lineages and hierarchy has not been reported. Recent reports indicate that prostate organoid cultures contain P-SCs derived from luminal cells (14,15). Together with urogenital mesenchymal cells, both the sphere-and organoid-forming cells can give rise to a glandular structure resembling the prostate in vivo (29).…”

“…For organoid culture in hepatocyte medium as described (14), the cells were resuspended in hepatocyte culture medium (designated as Medium H) that consisted of hepatocyte medium supplemented with 10 ng/ml of EGF (PeproTech), 10 m fasudil (Tocris), 1ϫ Glutamax (Gibco), 5% Matrigel (Corning), and 5% charcoal-stripped FBS (Gibco) that had been heat inactivated at 55°C for 1 h. Dissociated cells (10,000) were resuspended in Medium H and plated in ultralow attachment 96-well plates (Corning) in the presence of 100 nM dihydroxytesterone. Fresh Medium H (100 l) was added to the wells every other day.…”

“…Moreover, whether the prostasphere forming cells represent P-SCs is still controversial. Methods for growth of luminal-resident P-SCs in vitro remained problematic until two groups recently developed distinct culture systems (14,15). One group shows that both CARNs and normal prostate luminal epithelial cells can form prostate organoids and exhibit functional androgen receptors (AR) in culture.…”

Molecular Pathology of Patient Tumors, Patient-Derived Xenografts, and Cancer Cell Lines

“…The in vitro immortalized cell lines usually go through irreversible changes, or crisis, from their original tumors, resulting in loss of tissue specificity, while PDXs do not. It would be very interesting to compare the molecular pathology of human tumors with those of other new experimental systems that do not go through crisis and share histopathology features as in human tumors, such as organoid culture (21,22). If these systems do have similar molecular pathology, they could also function as predictive models, like PDXs.…”