Establishment of Three-Dimensional Bioprinted Bladder Cancer-on-a-Chip with a Microfluidic System Using Bacillus Calmette–Guérin

Abstract

Immunotherapy of bladder cancer is known to have favorable effects, although it is difficult to determine which patients will show a good response because of the different tumor microenvironments (TME). Here, we developed a bladder cancer-on-a-chip (BCOC) to mimic the TME using three-dimensional (3D) bioprinting and microfluidic technology. We fabricated a T24 and a 5637-cell line-based BCOC that also incorporated MRC-5, HUVEC, and THP-1 cells. We evaluated the effects of TME and assessed the immunologic reactions in response to different concentrations of Bacillus Calmette–Guérin (BCG) via live/dead assay and THP-1 monocytic migration, and concentrations of growth factors and cytokines. The results show that cell viability was maintained at 15% filling density in circle-shaped cell constructs at 20 μL/min microfluidic flow rate. A 3D co-culture increased the proliferation of BCOCs. We found that the appropriate time to evaluate the viability of BCOC, concentration of cytokines, and migration of monocytes was 6 h, 24 h, and three days after BGC treatment. Lastly, the immunotherapeutic effects of BCOC increased according to BCG dosage. To predict effects of immunotherapeutic agent in bladder cancer, we constructed a 3D bioprinted BCOC model. The BCOC was validated with BCG, which has been proven to be effective in the immunotherapy of bladder cancer.

The application of 3D bioprinting in urological diseases

“…The cellular viability of GelMA in bioprinting has been verified using numerous cell types, such as liver cells [ 98 ], nerves cells [ 99 ] and BMSCs [ 100 , 101 ]. Gelatin and GelMA have been 3D bioprinted for in vitro kidney models [ 102 , 103 ], urethral tubes [ 104 ], and bladder cancer models [ 105 , 106 ]. Similar methylation modifications were applied to other natural biomaterials to enhance their printability.…”

Immunotherapeutic effects of recombinant Bacillus Calmette–Guérin containing sic gene in ex vivo and in vivo bladder cancer models

“…Additionally, the T24 cell line was only used to evaluate the immunotherapeutic effects of the rBCG- sic . In a previous study, we investigated BCOC experiments with two different bladder cancer cell lines, T24 and 5637 [ 12 ]. We had reported better results in the T24 bladder cancer cell line at that time.…”

“…Complementing the previous study [ 12 ], we fabricated four BCOC-embedded microfluidic systems to enable efficient bladder cancer screening. Although the compositions of the BCOC models were similar, four stable experiments were performed using a single syringe pump ( Fig.…”

“…BCOC is an essential bladder cancer culture model for the development and establishment of an ex vivo cell-based system that can more realistically simulate cellular behavior in vivo [ 11 ]. Our BCOC is fabricated using 3D bioprinting technology with a microfluidic system, which can be used to create a bladder cancer-like environment that functions as a drug screening platform [ 12 ].…”

Perspectives for 3D-Bioprinting in Modeling of Tumor Immune Evasion

“…Kim et al fabricated a bioprinted bladder cancer model using T24 and 5637 bladder cancer cell lines alongside HUVEC and MRC-5 human lung fibroblasts to evaluate its effect on THP-1 human monocyte response to Bacillus Calmette-Guerin (BCG) [106]. They report a favorable secretion profile of THP-1 (IL6, IL12, IFN-γ and TNF-α), concurrent with clinically observed effects of BCG treatment.…”