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Evaluation of spheroid 3D culture methods to study a pancreatic neuroendocrine neoplasm cell line
journal contribution
posted on 2023-06-10, 01:43 authored by Giulia Bresciani, Leo J Hofland, Fadime Dogan, Georgios GiamasGeorgios Giamas, Teresa Gagliano, Maria Chiara ZatelliPancreatic Neuroendocrine Neoplasms (pNEN) are rare tumors which treatment still represent an important clinical problem, due to the paucity of medical treatments. Due to tumor complexity, techniques as 3D cultures are important to study drug activity in a more realistic model. 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. 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. 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 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. Moreover, we investigated how culture methods can influence experimental outcomes evaluating perimeter changes, cell viability and immunohistochemistry of spheroids treated with different Sunitinib concentrations. 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.
History
Publication status
- Published
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- Published version
Journal
Frontiers in EndocrinologyISSN
1664-2392Publisher
Frontiers MediaExternal DOI
Volume
10Page range
1-10Article number
a682Event location
SwitzerlandDepartment affiliated with
- Biochemistry Publications
Full text available
- Yes
Peer reviewed?
- Yes
Legacy Posted Date
2021-11-15First Open Access (FOA) Date
2021-11-15First Compliant Deposit (FCD) Date
2021-11-15Usage metrics
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