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In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival

Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental cells and signals. The lymph node (LN) is the critical site of in vivo CLL proliferation and development of resistance to both chemotherapy and targeted agents. We present a new model that incorporates key aspects of...

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Autores principales: Haselager, Marco V., van Driel, Bianca F., Perelaer, Eduard, de Rooij, Dennis, Lashgari, Danial, Loos, Remco, Kater, Arnon P., Moerland, Perry D., Eldering, Eric
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Lippincott Williams & Wilkins 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10448932/
https://www.ncbi.nlm.nih.gov/pubmed/37637994
http://dx.doi.org/10.1097/HS9.0000000000000938
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author Haselager, Marco V.
van Driel, Bianca F.
Perelaer, Eduard
de Rooij, Dennis
Lashgari, Danial
Loos, Remco
Kater, Arnon P.
Moerland, Perry D.
Eldering, Eric
author_facet Haselager, Marco V.
van Driel, Bianca F.
Perelaer, Eduard
de Rooij, Dennis
Lashgari, Danial
Loos, Remco
Kater, Arnon P.
Moerland, Perry D.
Eldering, Eric
author_sort Haselager, Marco V.
collection PubMed
description Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental cells and signals. The lymph node (LN) is the critical site of in vivo CLL proliferation and development of resistance to both chemotherapy and targeted agents. We present a new model that incorporates key aspects of the CLL LN, which enables investigation of CLL cells in the context of a protective niche. We describe a three-dimensional (3D) in vitro culture system using ultra-low attachment plates to create spheroids of CLL cells derived from peripheral blood. Starting from CLL:T cell ratios as observed in LN samples, CLL activation was induced by either direct stimulation and/or indirectly via T cells. Compared with two-dimensional cultures, 3D cultures promoted CLL proliferation in a T cell-dependent manner, and enabled expansion for up to 7 weeks, including the formation of follicle-like structures after several weeks of culture. This model enables high-throughput drug screening, of which we describe response to Btk inhibition, venetoclax resistance, and T cell-mediated cytotoxicity as examples. In summary, we present the first LN-mimicking in vitro 3D culture for primary CLL, which enables readouts such as real-time drug screens, kinetic growth assays, and spatial localization. This is the first in vitro CLL system that allows testing of response and resistance to venetoclax and Bruton's tyrosine kinase inhibitors in the context of the tumor microenvironment, thereby opening up new possibilities for clinically useful applications.
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spelling pubmed-104489322023-08-25 In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival Haselager, Marco V. van Driel, Bianca F. Perelaer, Eduard de Rooij, Dennis Lashgari, Danial Loos, Remco Kater, Arnon P. Moerland, Perry D. Eldering, Eric Hemasphere Article Chronic lymphocytic leukemia (CLL) cells are highly dependent on microenvironmental cells and signals. The lymph node (LN) is the critical site of in vivo CLL proliferation and development of resistance to both chemotherapy and targeted agents. We present a new model that incorporates key aspects of the CLL LN, which enables investigation of CLL cells in the context of a protective niche. We describe a three-dimensional (3D) in vitro culture system using ultra-low attachment plates to create spheroids of CLL cells derived from peripheral blood. Starting from CLL:T cell ratios as observed in LN samples, CLL activation was induced by either direct stimulation and/or indirectly via T cells. Compared with two-dimensional cultures, 3D cultures promoted CLL proliferation in a T cell-dependent manner, and enabled expansion for up to 7 weeks, including the formation of follicle-like structures after several weeks of culture. This model enables high-throughput drug screening, of which we describe response to Btk inhibition, venetoclax resistance, and T cell-mediated cytotoxicity as examples. In summary, we present the first LN-mimicking in vitro 3D culture for primary CLL, which enables readouts such as real-time drug screens, kinetic growth assays, and spatial localization. This is the first in vitro CLL system that allows testing of response and resistance to venetoclax and Bruton's tyrosine kinase inhibitors in the context of the tumor microenvironment, thereby opening up new possibilities for clinically useful applications. Lippincott Williams & Wilkins 2023-08-23 /pmc/articles/PMC10448932/ /pubmed/37637994 http://dx.doi.org/10.1097/HS9.0000000000000938 Text en Copyright © 2023 the Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the European Hematology Association. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the Creative Commons Attribution License 4.0 (CCBY) (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Article
Haselager, Marco V.
van Driel, Bianca F.
Perelaer, Eduard
de Rooij, Dennis
Lashgari, Danial
Loos, Remco
Kater, Arnon P.
Moerland, Perry D.
Eldering, Eric
In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title_full In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title_fullStr In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title_full_unstemmed In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title_short In Vitro 3D Spheroid Culture System Displays Sustained T Cell-dependent CLL Proliferation and Survival
title_sort in vitro 3d spheroid culture system displays sustained t cell-dependent cll proliferation and survival
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10448932/
https://www.ncbi.nlm.nih.gov/pubmed/37637994
http://dx.doi.org/10.1097/HS9.0000000000000938
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