Abstract Patient-derived cell (PDC) models are a tool that bridge the gap between conventional cancer lines that do not faithfully replicate clinical responses and costly and time-consuming patient derived xenograft (PDX) models. Imagen Therapeutics is establishing the world’s largest biobank of PDC models for both patient diagnostic (PredictRx) and biopharma drug development (PredictTx) studies that show a high degree of correlation between PDC and patient drug response. Through our research, we have established unique culture conditions that specifically select for cancer stem cells, and these have been characterized using both 2-D and 3-D short-term high content image (HCI)-based analysis. To better understand the genomic and phenotypic profiles of these models we analyzed Next Generation Sequencing data and phenotypic cell killing using a panel of FDA-approved agents. Our results showed that mRNA expression of several known cancer stem cell markers were upregulated in our PDCs in addition to multidrug resistance (MDR) genes. Drug resistance in the phenotypic assay was then correlated with stemness and MDR expression profiles. Finally, sequential PDCs (models developed from same patient at different stages in disease progression), in addition to PDCs developed from the same patient at multiple lesion sites, were compared in terms of genomic sequencing and drug responses. Together our data supports the premise that PDCs recapitulate the cancer stem cell population and are a useful platform with which to interrogate drug responses in a variety of different cancer types. Citation Format: Gareth J. Griffiths, Abbie Dodd, Olivia Nee, Tilly Thuringer, Subir Singh, Rachel Howard-Jones, Yuen Ngan Fan, Albert Bezman, Rachel Bell, Ido Sloma, Chris Noakes, Dominic I. James. Establishment and characterization of patient-derived cancer cell models for pharmaceutical drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 191.