Abstract Pancreatic ductal adenocarcinoma (PDAC) is known to be one of the deadliest cancers due to its late diagnosis and faster metastasis rate1. Thus, the time required to test the patient-derived cancer cells against different therapeutics becomes extremely crucial. Gene expression analysis using real-time PCR is vital, typically when designing patient-specific therapeutics. 2D tumor models do not represent the true picture of the tumor in vivo. In order to perform these gene expression studies, robust 3D cancer models are required2. The traditional techniques of generating cancer spheroids using U-bottom wells and hanging drop methods are effective but are limited by scalability as well as the limitations associated with the traditional organoid architecture. Gene expression studies of spheroids under different combinations of drug treatment thus requires many well defined and easy to handle spheroids. Here, we are trying to perform real-time PCR studies to access the gene expression changes on MIA PaCa-2 microgel based spheroids generated using co-axial droplet microfluidics with or without stromal cells. This 3D platform will enable gene expression studies in miniature co-culture tumor models. In addition, PCR and single-cell sequencing analyses will define the gene expression changes in the recently described conditionally reprogramed, patient-derived, primary PDAC cultures3. The establishment and analysis of this 3D model is fast, reproducible and can be easily scaled to enable high-throughput screening, thereby providing an opportunity to design patient-specific precision medicine. The research leading to these results received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement No 759959, ERC-StG “INTERCELLMED”). References: (1) Adamska, A.; Domenichini, A.; Falasca, M. Pancreatic Ductal Adenocarcinoma: Current and Evolving Therapies. Int. J. Mol. Sci. 2017, 18 (7). (2) Moreira, L.; Bakir, B.; Chatterji, P.; Dantes, Z.; Reichert, M.; Rustgi, A. K. Pancreas 3D Organoids: Current and Future Aspects as a Research Platform for Personalized Medicine in Pancreatic Cancer. Cell. Mol. Gastroenterol. Hepatol. 2018, 5 (3), 289-298. (3) Parasido, E.; Avetian, G. S.; Naeem, A.; Graham, G.; Pishvaian, M.; Glasgow, E.; Mudambi, S.; Lee, Y.; Ihemelandu, C.; Choudhry, M.; et al. The Sustained Induction of C-MYC Drives Nab-Paclitaxel Resistance in Primary Pancreatic Ductal Carcinoma Cells. Mol. Cancer Res. 2019, 17 (9), 1815-1827. Citation Format: Saumya Prasad, Anil Chandra, Enza Lonardo, Erika Parasido, Christopher Albanese, Loretta L. del Mercato. Gene expression studies using microgel embedded pancreatic cancer spheroids [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1577.