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pNEN diagnosis is mostly influenced by hormonal hypersecretion that usually leads to early diagnosis due to the manifestation of clinical symptoms ( 3). pNEN represent 7% of all NEN and ~2% of all pancreatic neoplasms with an incidence of 1–2 per 100,000 persons per year ( 2). Pancreatic Neuroendocrine Neoplasms (pNEN) are malignancies arising from the Islets of Langerhans and, therefore, are different from the more frequently occurring exocrine pancreas cancer ( 1). Results showed that each method has weak and strong points but, considering the easiness of spheroids maintenance and reproducibility results, ULA plates method appears to be the best approach to culture BON1 spheroids and, therefore, to study pNEN. Moreover, we investigated how culture methods can influence experimental outcomes evaluating perimeter changes, cell viability and immunohistochemistry of spheroids treated with different Sunitinib concentrations. The evaluated parameters during the study were: cell seeding, easiness in spheroids formation, morphology, culture maintenance, medium change, spheroids monitoring, picture quality, spheroid perimeter measurement reproducibility error, possibility to perform assays into the seeding plate, overall time of the experiment. The investigated methods are: (1) 96-well hanging drop plates (HD plates), (2) 24-well plates with a cell-repellent surface, and (3) ultra-low attachment 96-well plates with clear round bottom (ULA plates). The BON1 cell line was used as a pNEN model and the well-known Receptor Tyrosine Kinase inhibitor Sunitinib was used in order to better investigate the different features of each method. This study aims to compare three different 3D culture methods in order to understand which one can be considered the best option in terms of experimental easiness and reproducibility in studying the efficacy of a target drug on pNEN. Due to tumor complexity, techniques as 3D cultures are important to study drug activity in a more realistic model. Pancreatic Neuroendocrine Neoplasms (pNEN) are rare tumors which treatment still represent an important clinical problem, due to the paucity of medical treatments. 3Department of Biochemistry and Biomedicine, School of Life Science, University of Sussex, Brighton, United Kingdom.2Department of Internal Medicine, Erasmus Medical Center, Rotterdam, Netherlands.
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Hofland 2, Fadime Dogan 2, Georgios Giamas 3, Teresa Gagliano 3 and Maria Chiara Zatelli 1 *