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Table 1 Culture methods used to generate 3D preclinical models of prostate cancer

From: Patient derived organoids in prostate cancer: improving therapeutic efficacy in precision medicine

Starting material Media Serum Growth factors Scaffold Basement Membrane Equivalent (BME) Culture method Features Ref no.
Scaffold-free 3D cultures
 PCa cell lines (LNCaP, PC3 and DU145) RPMI 1640 (LNCaP) and DMEM (PC3 and DU145) 10% FBS None None Liquid overlay method Single aggregates are formed in the well using 1% Seaplaque-agarose with gentle agitation [10]
 PCa cell line (LNCaP) and human osteoblast cell line (MG63) in co-culture T-medium 10% FBS None None Rotary wall vessel (RWV) culture Microgravity conditions with perpetual fluid rotation [11]
 PCa cell line (LNCaP) DMEM 10% FBS None None Hanging drop culture Non-adherent conditions are created using hanging drop plates sealed with agarose [12, 17]
 PDX from primary PCa ADMEM/F12 with HEPES, GlutaMAX and Primocin 5% FBS Y-27632, Nicotinamide, Noggin, Rspondin, N-acetyl-cysteine, DHT, Wnt3a, HGF, EGF, FGF10, FGF2, PGE2, SB202190 (P38 MAP Kinase
inhibitor), and A83–01 (TGF-b
pathway inhibitor)
None None Organoids grown in suspension scaffold-free conditions, with no requirement for ECM support [18]
Matrigel-based 3D cultures
 Non-malignant human prostate tissue Keratinocyte serum-free medium (KSFM) None EGF and BPE 50% Matrigel Matrigel embedding Patient stromal cells were cocultured with prostate cells in transwell inserts [13]
 Malignant and nonmalignant human prostate tissue and immortalized cell lines KSFM None EGF, BPE, and R1881 (synthetic androgen agonist) 2% matrigel Matrigel suspension Matrigel enabled differentaition of non-malignant prostate epithelial cells and cocultured with irradiated fibroblasts, which were reprogrammed to stem-like cells [14, 16]
 Human prostates from radical prostatectomy and murine prostates ADMEM F12 with B27, HEPES, Glutamax, and Penicillin/Streptomycin None EGF, R-spondin 1, Noggin, A83–01, DHT, FGF10, FGF2, Prostaglandin E2, SB202190, Nicotinamide 100% Matrigel Matrigel embedding Optimized conditions for continuous propagation of normal human basal and luminal prostate epithelial cells [19,20,21,22,23,24,25,26,27]
 Human prostate from radical prostatectomy, PCa cell line (VCaP) and murine prostate Hepatocyte medium with Glutamax and Penicillin/Streptomycin 5% charcoalstripped FBS EGF, Y-27632, DHT 5% Matrigel (60% matrigel for drug treatment) Matrigel suspension Matrigel embedding for drug treatments) Bipotential Luminal progenitor derived organoids were cultured for upto 13 passages [28, 29]
 PCa cell lines KSFM for RWPE1 and RPMI-1640 for PCa cell lines 2% FBS None 50% Matrigel (bottom layer) 25% matrigel (top layer) Matrigel Sandwich A comprehensive panel of miniaturized prostate cell culture derived from cell lines was developed [30]
 Patient cells from radical prostatecomy KSFM 5% charcoalstripped FBS DHT 50% Matrigel (bottom layer) 33% matrigel (top layer) Matrigel sandwich Co-culture with human primary prostate stromal cells improved epithelial organoid viability [31]
Synthetic scaffold-based 3D cultures
 Cell lines (RWPE-1, RWPE-2, and LNCaP) KSFM None EGF and BPE Polydimethylsiloxa ne (PDMS) PDMS Microwell platform Microwell arrays enabled formation of prostate micro aggregates of defined size suitable for HTS [32, 33]
 MDA PCa 118b PDX ADMEM/F-12 and alpha- MEM medium 2% FBS None Thiolated SH: acrylate-PEGGRGDS: acrylate- PEG-PQ-PEGacrylate (4:1:1) Synthetic organoid culture PDX tumor cells and osteoblasts were encapsulated together in a 3D hyaluronan (HA) hydrogel. Tumor cell–hydrogel mixture encapsulated in custom-made PDMS molds. [34]
 PCa patient biopsies ADMEM None B27, N-Acetylcysteine, EGF, FGF-10, FGF-basic, A-83-01, SB202190, Nicotinamide DHT, PGE2, Noggin conditioned media and R-spondin conditioned media PEG-4MAL macromer and cell cross linker at 1:1 ratio Synthetic organoid culture Organoids were cultured in synthetic hydrogels that recapitulate PCa ECM [35]