Abstract

Two-dimensional (2D) cell culture is a valuable method for cell-based research but can provide unpredictable, misleading data about in vivo responses. In this study, we created a three-dimensional (3D) cell culture environment to mimic tumor characteristics and cell-cell interactions to better characterize the tumor formation response to chemotherapy. We fabricated the 3D cell culture samples using a 3D cell bio printer and the bladder cancer cell line 5637. T24 cells were used for 2D cell culture. Then, rapamycin and Bacillus Calmette-Guérin (BCG) were used to examine their cancer inhibition effects using the two bladder cancer cell lines. Cell-cell interaction was measured by measuring e-cadherin and n-cadherin secreted via the epithelial-mesenchymal transition (EMT). We constructed a 3D cell scaffold using gelatin methacryloyl (GelMA) and compared cell survival in 3D and 2D cell cultures. 3D cell cultures showed higher cancer cell proliferation rates than 2D cell cultures, and the 3D cell culture environment showed higher cell-to-cell interactions through the secretion of E-cadherin and N-cadherin. Assessment of the effects of drugs for bladder cancer such as rapamycin and BCG showed that the effect in the 2D cell culture environment was more exaggerated than that in the 3D cell culture environment. We fabricated 3D scaffolds with bladder cancer cells using a 3D bio printer, and the 3D scaffolds were similar to bladder cancer tissue. This technique can be used to create a cancer cell-like environment for a drug screening platform.

Highlights

  • The cell culture system was an essential method that is often used in basic and clinical in vivo studies

  • Assessment of the effects of drugs for bladder cancer such as rapamycin and Bacillus Calmette-Guerin (BCG) showed that the effect in the 2D cell culture environment was more exaggerated than that in the 3D cell culture environment

  • We fabricated 3D scaffolds with bladder cancer cells using a 3D bio printer, and the 3D scaffolds were similar to bladder cancer tissue

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Summary

Introduction

The cell culture system was an essential method that is often used in basic and clinical in vivo studies. Cell culture is an important technique in the drug discovery process, providing a simple, fast, and cost-effective way to reduce animal testing.[1] Two-dimensional (2D) cell culture is a valuable method for cell-based research but has limitations.[2] Almost all cells in the in vivo environment are surrounded by extracellular matrix (ECM) and other cells. 2D cell culture sometimes provides unpredictable data that can be misleading regarding the in vivo response.[3] Currently, standard procedures for compound screening in new drug development begin with 2D cell culture-based testing and move to animal model testing and clinical trials. We hypothesized that the effect of rapamycin (mTOR inhibitor) and BCG in the 3D cell culture system would be less than that observed in 2D, indicating that 3D cell culture is a more suitable in vivo model

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