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Background: Malignant pleural mesothelioma (MPM) is an incurable and aggressive malignancy, which is typically presented with malignant pleural effusion (MPE), bearing severely comprised quality of life and short survival. Since management of MPE remains symptomatic and targeted MPM treatments are not available the development of translational models to improve patients’ stratification are desperately needed. Aim: To establish a panel of patient derived MPM cell lines which can serve as a faithful model for the discovery of novel mutations and the development of targeted treatments. Methods: MPE samples from MPM patients were collected and cultured to establish a pure cancer cell population. To cross-examine the genomic background of the MPM cells between different passages, a proportion of the cells was kept from each passage. Results: We established a panel of 18 primary personalised mesothelioma cell (PMC) lines. Phenotypically PMC lines exhibited pleomorphic, atypical and often multiple nucleoli. In vitro, the cells were immortal and able to form colonies and tumour-spheres. A high-throughput drug screening assay was used to identify the cluster of the most potent anticancer drugs, demonstrating a heterogeneous response for each PMC line. Genome profiling revealed no major differences for important cancer genes among the passages. T cells from the MPEs were isolated, cultured and examined regarding their cellular and molecular phenotype. Conclusions: We developed a methodology to establish patient derived MPM cell lines which could potentially be used as tools to refine patients’ management.