Fiche publication
Date publication
avril 2026
Journal
Journal of visualized experiments : JoVE
Auteurs
Membres identifiés du Cancéropôle Est :
Dr AUGER Cyril
Tous les auteurs :
Saidi A, Tollard C, Liu L, El Khallouqi Y, Sévigny J, Auger C, Kauffenstein G, Ghazouani FE, Kessler L, Toti F
Lien Pubmed
Résumé
This protocol details an optimized method for the production of small stable spheroids, their culture, and 3D imaging, for the study of the endothelial and insulin-producing β cells interactions in a 3D model of pancreatic islets. The 150-200 µm spheroids, mirroring the lowest range of islet sizes, were prepared from a selected ratio combining 1 intra-islet endothelial cells (MS-1 cells) to 20 insulin-secreting cells (β-TC-6). Staining, clearing, and mounting challenges of small spheroids and their tackling by employing low-melting point agarose and the CUBIC clearing technique are detailed, as well as key points for an efficient analysis of the 3D structure with different probes. Data indicate that NTPDASE-ectonucleotidase 3 does not colocalize with insulin in the spheroid model, suggesting varying maturity and functional levels of β-TC6 and that the complete procedure can also be applied to isolated pancreatic islets, with clear probing of intra-islet vessels. These findings underscore the effectiveness of the 3D imaging protocol in revealing complex pancreatic cell organization and interactions within the islet model.
Mots clés
Insulin-Secreting Cells, cytology, Animals, Endothelial Cells, cytology, Islets of Langerhans, cytology, Spheroids, Cellular, cytology, Mice, Imaging, Three-Dimensional, methods, Cell Communication, physiology, Cell Culture Techniques, Three Dimensional, methods
Référence
J Vis Exp. 2026 04 17;(230):
