Establishment of a three-dimensional (3D) lung cancer model for translational research

Abstract

In the past, different spheroid-, organotypic-, and three-dimensional (3D) bioprinting lung cancer models were established for in vitro drug testing and personalized medicine. These tissue models cannot depict the tumor microenvironment (TME) and therefore research addressing tumor cell-TME interactions is limited. To overcome this hurdle, we applied patient-derived lung tumor samples to establish new in vitro 3D models with three different culture conditions: static, dynamic, and co-cultivation. All three culture conditions afforded tissue culture for up to 28 days. The success rates of the static 3D lung cancer model was significantly higher than the corresponding 2D model. Our tissue models were characterized by hematoxylin eosin staining and immunofluorescence staining. We found tumor cells that were positive for specific lung cancer markers (TTF-1 and p40/p63), cancer associated fibroblasts (CAFs) positive for α-SMA and MCT4, and extracellular matrix for fibronectin (FN). Using DAPI staining, we determined the cell density of the static 3D model which increased from day 21 to day 28. Apoptosis measured through the M30 ELISA decreased from day 21 to day 28. 3D models which were co-cultured with NIH-3T3 cells, however, showed lower apoptosis in general. In addition, we performed preliminary tests culturing the cells on the SISmuc matrix for replacement materials of our 3D model. Taken together, a novel in vitro 3D lung cancer model was established, which simulated the TME for 28 days and possessed a structural complexity.