Three-dimensional decellularized amnion membrane scaffold as a novel tool for cancer research; cell behavior, drug resistance and cancer stem cell content

Cancer is the second leading cause of human death. Therefore, comprehensive research and the appropriate tools are needed in this field. Animal models and cell culture studies are the most important preclinical tools in cancer research. In 2D cell culture models, cells are forced to grow in a 2D environment, which differs from their natural physiology. Recently, 3D cell culture models were developed to fill the gap between 2D cell culture and animal models.

Human amniotic membranes were obtained, decellularized, characterized and used as a natural 3D scaffold to investigate cancer cell behavior in 2D compared to 3D conditions. Time-lapse imaging of cells was used, and cell proliferation, velocity and migration were evaluated. Cisplatin was applied in 2D and 3D conditions, followed by evaluation of viability, apoptosis and cancer stem cell proteins by flow cytometry and western blot analysis.

The results showed that in the decellularized amnion membrane (DAM) scaffold most cells did not spread and remain rounded and then penetrated into the scaffold with no cytotoxicity. Significant differences in migration, velocity, morphology and proliferation of cancer cells were observed between the 3D DAM scaffold and 2D model. Furthermore, the cells in the 3D DAM scaffold showed much more resistance to apoptosis and higher CSC content.

In conclusion, considering the effect of the 3D DAM scaffold in cell behavior, apoptosis resistance and CSC content as well as the short processing time for decellularizing the AM, it appears that the 3D DAM scaffold offers an appropriate tool for in vitro cancer research.