Table 1 Summary of studies used three-dimensional (3D) culture models in bladder cancer

3D bio-printed and 2D cell cultures of T24 and 5637 cells

T24 and 5637 were cultured on a synthetic 3D scaffold.

The cell survival rates in the 3D and 2D cultures and sensitivity of cells to rapamycin and Bacillus Calmette-Guérin (BCG)

Cells of 3D cultures demonstrated higher proliferation rates and more exaggerated response to rapamycin and BCG than those of 2D cultures

[114]

Bladder and prostate cancer cell lines

Spheroids were generated from T24 and SV-HUC-1.

The cytotoxic effect of ciprofloxacin and levofloxacin on cell lines during culture

Both drugs exhibited a toxic effect on the tested cell lines (↑ apoptosis; ↓ S phase cell proliferation).

[115]

Tumor cells from BC patients and BC cell lines (RT4, UM-UC-3, and HT1376)

Microtumors were created using a self-assembly process.

The gene expression profiles of cells of the 3D microtumors and those of traditional cultures

A more invasive phenotype was observed in 3D microtumors that was associated with upregulated expression of Delta-like ligand 4 (DLL4)

[116]

Organoids of human BC cell lines and primary cancer cells

Primary cell organoids (BCa #01)

The effect of Wnt/β-catenin pathway activation, using CHIR99021, on cancer cell proliferation

Wnt/β-catenin activation increased proliferation of BC cells grown in 3D cultures but not in conventional adherent systems

[117]

Human urothelial cancer of the bladder (HUCB)

3D co-cultured spheres of HUCB cells and tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs).

The paracrine effect of TAMs/CAFs on tumor microenvironment

3D co-culture of HUCB cells and TAMs/CAFs increased CXCL1 production in culture with subsequent increase in cell-to-cell interaction among cancer cells and TAMs/CAFs

[118]

3D-spheroids of BC cell lines RT4 and 5637

RT4 and 5637 spheroids were prepared using the aggregation-based method. 2 × 10⁵ or 1,000 cells (respectively) were seeded in 6- or 96-well U-bottom plates coated with poly-HEMA.

Protein expression of the luminal markers peroxisome proliferator activated receptor γ (PPARγ) and forkhead box A1 (FOXA1) in cancer spheroids

PPARγ and FOXA1 proteins were expressed to a lesser extent in cancer spheroids than in cells grown in 2D cultures.

[119]

3D-spheroids of human BC primary cells

Biopsies from bladder tumors were fragmented and allowed to form 3D spheroids.

Sensitivity of the cancer cells to the chemotherapeutic agents mitomycin C, thiotepa, epirubicin, and adriamycin

Mitomycin C achieved the best results with mean sensitivity of 50%, followed by thiotepa (37%), epirubicin (7%), and adriamycin  (3%).

[120]

Prostate and bladder cancer cell lines

5637 and T24/TSU-Pr1 cell lines were pelleted and resuspended into 50 mL Bioreactor tubes at density of 100,000 cells/mL.

Comprehensive metabolomic analysis of cells of 3D and 2D cultures

The cells of 3D culture had significantly higher metabolites levels than those of the 2D culture

[121]

BC cell lines (RT4 and PDX)

3D spheroids of RT4 cells were generated using 96-well micro honeycomb plates (1 × 10⁴ cells/well with 2% of Matrigel); 3D spheroids of PDX cells were generated using 96-well low attachment plates (3.9 × 10⁴ cells/well, without Matrigel)

Chemosensitizing effect of glycoalkaloids with cisplatin in RT4 and PDX cells using 2D and 3D cell culture models.

Significantly higher IC₅₀ values in cells of 3D cultures than those of 2D monolayers of both RT4 and PDX.

[122]