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Article

3D Model Characterization by 2D and 3D Imaging in t(14;18)-Positive B-NHL: Perspectives for In Vitro Drug Screens in Follicular Lymphoma

1
Centre de Recherches en Cancérologie de Toulouse, INSERM UMR1037, CEDEX 1, 31037 Toulouse, France
2
Université Toulouse III Paul-Sabatier, CEDEX 9, 31062 Toulouse, France
3
ERL 5294 CNRS, CEDEX 4, 31055 Toulouse, France
4
Institut Universitaire du Cancer-Oncopole de Toulouse, CEDEX 9, 31059 Toulouse, France
5
Laboratoire d’Excellence ‘TOUCAN-2’, CEDEX 1, 31037 Toulouse, France
6
Institut Carnot Lymphome CALYM, 69495 Pierre-Bénite, France
7
Department of Pathology, Institut Universitaire du Cancer de Toulouse, CEDEX 9, 31059 Toulouse, France
8
Department of Hemato-Oncology, IDIBAPS, 08036 Barcelona, Spain
9
Centro de Investigación Biomédica en Red-Oncología (CIBERONC), 28029 Madrid, Spain
10
IMACTIV-3D, 1 Place Pierre Potier, 31106 Toulouse, France
11
CHU Dijon, Hématologie clinique, Hôpital François Mitterand, 21000 Dijon, France
12
RESTORE Research Center, Université de Toulouse, INSERM, CNRS, EFS, ENVT, 31100 Toulouse, France
13
Department of Hematology, Institut Universitaire du Cancer de Toulouse, CEDEX 9, 31059 Toulouse, France
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Ellen Leich
Cancers 2021, 13(7), 1490; https://doi.org/10.3390/cancers13071490
Received: 29 January 2021 / Revised: 5 March 2021 / Accepted: 6 March 2021 / Published: 24 March 2021
(This article belongs to the Section Methods and Technologies Development)
Follicular lymphoma is an indolent B cell lymphoproliferative disorder of transformed follicular center B cells, which accounts for 20–30 percent of all non-Hodgkin lymphoma (NHL) cases. Although huge efforts have been made in the last 10 years, this pathology is still considered as incurable, leaving open the discovery and testing of new therapeutic targets requiring relevant preclinical models. Here, we report a realistic 3D model of t (14;18)-positive B-NHL cell culture (ultra-low attachment (ULA)-multicellular aggregates of lymphoma cells (MALC)), which monitored by state-of-the-art 2D and 3D imaging, allows more robust drug testing.
Follicular lymphoma (FL) is an indolent B cell lymphoproliferative disorder of transformed follicular center B cells, which accounts for 20–30 percent of all non-Hodgkin lymphoma (NHL) cases. Great advances have been made to identify the most relevant targets for precision therapy. However, no relevant models for in vitro studies have been developed or characterized in depth. To this purpose, we generated a 3D cell model from t(14;18)-positive B-NHL cell lines cultured in ultra-low attachment 96-well plates. Morphological features and cell growth behavior were evaluated by classical microscopy (2D imaging) and response to treatment with different drugs was evaluated by a high-content analysis system to determine the robustness of the model. We show that the ultra-low attachment (ULA) method allows the development of regular, spherical and viable ULA-multicellular aggregates of lymphoma cells (MALC). However, discrepancies in the results obtained after 2D imaging analyses on drug-treated ULA-MALC prompted us to develop 3D imaging and specific analyses. We show by using light sheet microscopy and specifically developed 3D imaging algorithms that 3D imaging and dedicated analyses are necessary to characterize morphological properties of 3D models and drug effects. This study proposes a new method, but also imaging tools and informatic solutions, developed for FL necessary for future preclinical studies. View Full-Text
Keywords: follicular lymphoma; 3D model; spheroid; drug testing; 2D imaging; SPIM follicular lymphoma; 3D model; spheroid; drug testing; 2D imaging; SPIM
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MDPI and ACS Style

Gava, F.; Faria, C.; Gravelle, P.; Valero, J.G.; Dobaño-López, C.; Morin, R.; Norlund, M.; Gomes, A.; Lagarde, J.-M.; Rossi, C.; Bordenave, J.; Pieruccioni, L.; Rouquette, J.; Matas-Céspedes, A.; Fournié, J.-J.; Ysebaert, L.; Laurent, C.; Pérez-Galán, P.; Bezombes, C. 3D Model Characterization by 2D and 3D Imaging in t(14;18)-Positive B-NHL: Perspectives for In Vitro Drug Screens in Follicular Lymphoma. Cancers 2021, 13, 1490. https://doi.org/10.3390/cancers13071490

AMA Style

Gava F, Faria C, Gravelle P, Valero JG, Dobaño-López C, Morin R, Norlund M, Gomes A, Lagarde J-M, Rossi C, Bordenave J, Pieruccioni L, Rouquette J, Matas-Céspedes A, Fournié J-J, Ysebaert L, Laurent C, Pérez-Galán P, Bezombes C. 3D Model Characterization by 2D and 3D Imaging in t(14;18)-Positive B-NHL: Perspectives for In Vitro Drug Screens in Follicular Lymphoma. Cancers. 2021; 13(7):1490. https://doi.org/10.3390/cancers13071490

Chicago/Turabian Style

Gava, Fabien, Carla Faria, Pauline Gravelle, Juan G. Valero, Cèlia Dobaño-López, Renaud Morin, Marine Norlund, Aurélie Gomes, Jean-Michel Lagarde, Cédric Rossi, Julie Bordenave, Laetitia Pieruccioni, Jacques Rouquette, Alba Matas-Céspedes, Jean-Jacques Fournié, Loïc Ysebaert, Camille Laurent, Patricia Pérez-Galán, and Christine Bezombes. 2021. "3D Model Characterization by 2D and 3D Imaging in t(14;18)-Positive B-NHL: Perspectives for In Vitro Drug Screens in Follicular Lymphoma" Cancers 13, no. 7: 1490. https://doi.org/10.3390/cancers13071490

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