An Optimized Method to Culture Human Primary Lung Tumor Cell Spheroids

Simple Summary Lung cancer, responsible for nearly 20% of global cancer-related deaths annually, is the leading cause of cancer mortality. Traditional treatments, like chemotherapy and radiation, are not always effective or appropriate due to the heterogeneity of lung cancer across patients. To address this challenge, our research introduces an approach to develop patient-derived spheroids (PDS) that are cultivated using cells from patient tumors and adjacent healthy tissue. These PDS allow us to mimic the unique microenvironment of patients, introducing a platform to assess personalized responses to drug treatments. We used this system to characterize patient cell populations, evaluate gene expression, and assess the sensitivity of PDS to specific drugs. By introducing a patient-specific platform to test drug sensitivity, our study holds the potential to enhance the efficacy of lung cancer treatments, paving the way for individualized and more effective lung cancer therapies in the future.Abstract Lung cancer is the leading cause of cancer mortality worldwide, with a median survival rate at 5 years of less than 20%. While molecular mapping aids in selecting appropriate therapies, it cannot predict personalized treatment response and long-term efficacy. For addressing these challenges, there is a great need for functional tests. Within this context, we developed patient-derived spheroids (PDS) from tumor and adjacent normal tissue to biomimic the respective tissue for assessing the personalized drug treatment response in vitro. Surgically resected lung specimens were used to generate spheroids using a two-step culture procedure. Flow cytometry and immune staining enabled the characterization of different cell populations resulting from the lung samples. PDS phenotype, cell proliferation and apoptosis were evaluated. Differential gene expression between tumor and adjacent normal tissue was analyzed via RT-qPCR. PDS drug sensitivity was assessed using a cell metabolic assay in response to two chemotherapeutic drug combinations. Cellular and molecular analysis revealed the proportion of epithelial cells, fibroblasts, and immune cells in the patients' tissue samples. Subsequently, PDS models from tumor and normal lung were successfully established using the expanded epithelial cells. As a proof of concept, an analysis of the drug treatment using PDS of lung adenoid cystic carcinoma exhibited a dose-dependent effect in response to cisplatin/etoposide and cisplatin/paclitaxel. Our spheroid model of both tumor and non-tumor lung cells holds great promise for enhancing the treatment efficacy in the cancer patients.