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

 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]

 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]

 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]