3D Scaffold-Based Culture System Enhances Preclinical\nEvaluation of Natural Killer Cell Therapy in A549 Lung Cancer Cells

Cell-based immunotherapies have emerged\nas promising cancer treatment\nmodalities, demonstrating remarkable clinical efficacy. As interest\nin applying immune cell-based therapies to solid tumors has gained\nmomentum, experimental models that enable long-term monitoring and\nmimic clinical administration are increasingly necessary. This study\nexplores the potential of scaffold-based cell culture technologies,\nspecifically three-dimensional (3D) extracellular matrix (ECM)-like\nframeworks, as promising solutions. These frameworks facilitate unhindered\nimmune cell growth and enable continuous cancer cell culture. The\nthree-dimensional (3D) cell culture model was developed using tailored\nscaffolds for natural killer (NK) cell culture. Within this framework,\nA549 lung cancer cells were cocultured with NK cells, allowing real-time\nmonitoring for up to 28 days. The expression of critical markers associated\nwith anticancer drug resistance and epithelial-mesenchymal transition\n(EMT) was evaluated in cancer cells within this 3D culture context.\nCompared to conventional 2D monolayer cultures, this 3D scaffold-based\nculture revealed that solid tumor cells, specifically A549 cells,\nexhibited heightened resistance to anticancer drugs. Additionally,\nthe 3D culture environment upregulated the expression of EMT markers\nnamely vimentin, N-cadherin, and fibronectin, while NK and zEGFR-CAR-NK\ncells displayed anticancer effects. In the two-dimensional (2D) coculture,\nonly zEGFR-CAR-NK cells exhibited such effects in the 3D coculture\nsystem, highlighting an intriguing inconsistency with the 2D culture\nmodel, further confirmed by <i>in vivo</i> experiments.\nThis <i>in vitro</i> 3D cell culture model reliably predicts\noutcomes in NK immunotherapy experiments. Thus, it represents a valuable\ntool for investigating drug resistance mechanisms and assessing the\nefficacy of immune cell-based therapies. By bridging the gap between <i>in vitro</i> and <i>in vivo</i> investigations, this\nmodel effectively translates potential treatments into animal models\nand facilitates rigorous preclinical evaluations.