Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation

Summary: Hydrogel-based 3D culture systems are increasingly used for preclinical evaluation of cell-based immunotherapies, including chimeric antigen receptor T (CAR-T) cells. However, hydrogel properties can influence T cell behavior, potentially affecting interpretation of immunotherapy studies. W...

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Main Authors: Sonia Aristin Revilla, Alessandro Cutilli, Eugenia Cambiaso, Dedeke Rockx-Brouwer, Cynthia Lisanne Frederiks, Marc Falandt, Riccardo Levato, Onno Kranenburg, Caroline A. Lindemans, Paul James Coffer, Victor Peperzak, Enric Mocholi, Marta Cuenca
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:iScience
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589004225014956
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author Sonia Aristin Revilla
Alessandro Cutilli
Eugenia Cambiaso
Dedeke Rockx-Brouwer
Cynthia Lisanne Frederiks
Marc Falandt
Riccardo Levato
Onno Kranenburg
Caroline A. Lindemans
Paul James Coffer
Victor Peperzak
Enric Mocholi
Marta Cuenca
author_facet Sonia Aristin Revilla
Alessandro Cutilli
Eugenia Cambiaso
Dedeke Rockx-Brouwer
Cynthia Lisanne Frederiks
Marc Falandt
Riccardo Levato
Onno Kranenburg
Caroline A. Lindemans
Paul James Coffer
Victor Peperzak
Enric Mocholi
Marta Cuenca
author_sort Sonia Aristin Revilla
collection DOAJ
description Summary: Hydrogel-based 3D culture systems are increasingly used for preclinical evaluation of cell-based immunotherapies, including chimeric antigen receptor T (CAR-T) cells. However, hydrogel properties can influence T cell behavior, potentially affecting interpretation of immunotherapy studies. We assessed CD4+ T and CAR-T cell responses in two chemically undefined matrices—Matrigel and basement membrane extract (BME)— and in a synthetic nanofibrillar cellulose (NFC) hydrogel. Although NFC was mechanically stiffer, T cell activation and proliferation were higher in NFC than in Matrigel or BME. Murine CD4+ T cells acquired a regulatory phenotype in Matrigel and BME but not in NFC. Similarly, CAR-T cell function was reduced in Matrigel and BME but maintained in NFC. These findings underscore how matrix composition can shape T cell responses in 3D culture. NFC provides a chemically defined alternative that preserves T cell activity, supporting its use in more accurate preclinical testing of immunotherapies.
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spelling doaj-art-788024e355194bbca26ffddb6cf3ac732025-08-20T04:01:03ZengElsevieriScience2589-00422025-09-0128911323410.1016/j.isci.2025.113234Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activationSonia Aristin Revilla0Alessandro Cutilli1Eugenia Cambiaso2Dedeke Rockx-Brouwer3Cynthia Lisanne Frederiks4Marc Falandt5Riccardo Levato6Onno Kranenburg7Caroline A. Lindemans8Paul James Coffer9Victor Peperzak10Enric Mocholi11Marta Cuenca12Center for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Regenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Laboratory Translational Oncology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsCenter for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Regenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the NetherlandsCenter for Translational Immunology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsCenter for Translational Immunology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsCenter for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Regenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Laboratory Translational Oncology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsRegenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3584 CM Utrecht, the NetherlandsRegenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Department of Clinical Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3584 CM Utrecht, the Netherlands; Department of Orthopaedics, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsLaboratory Translational Oncology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsRegenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Princess Máxima Center for Pediatric Oncology, Heidelberglaan 25, 3584 CS Utrecht, the Netherlands; Department of Pediatrics, Wilhelmina Children’s Hospital University Medical Center Utrecht, Utrecht, the NetherlandsCenter for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Regenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the NetherlandsCenter for Translational Immunology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the NetherlandsCenter for Molecular Medicine, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Regenerative Medicine Center, University Medical Center Utrecht, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands; Corresponding authorCenter for Translational Immunology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, the Netherlands; Corresponding authorSummary: Hydrogel-based 3D culture systems are increasingly used for preclinical evaluation of cell-based immunotherapies, including chimeric antigen receptor T (CAR-T) cells. However, hydrogel properties can influence T cell behavior, potentially affecting interpretation of immunotherapy studies. We assessed CD4+ T and CAR-T cell responses in two chemically undefined matrices—Matrigel and basement membrane extract (BME)— and in a synthetic nanofibrillar cellulose (NFC) hydrogel. Although NFC was mechanically stiffer, T cell activation and proliferation were higher in NFC than in Matrigel or BME. Murine CD4+ T cells acquired a regulatory phenotype in Matrigel and BME but not in NFC. Similarly, CAR-T cell function was reduced in Matrigel and BME but maintained in NFC. These findings underscore how matrix composition can shape T cell responses in 3D culture. NFC provides a chemically defined alternative that preserves T cell activity, supporting its use in more accurate preclinical testing of immunotherapies.http://www.sciencedirect.com/science/article/pii/S2589004225014956Biological sciencesImmune responseCell biologyMaterials scienceBiomaterials
spellingShingle Sonia Aristin Revilla
Alessandro Cutilli
Eugenia Cambiaso
Dedeke Rockx-Brouwer
Cynthia Lisanne Frederiks
Marc Falandt
Riccardo Levato
Onno Kranenburg
Caroline A. Lindemans
Paul James Coffer
Victor Peperzak
Enric Mocholi
Marta Cuenca
Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
iScience
Biological sciences
Immune response
Cell biology
Materials science
Biomaterials
title Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
title_full Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
title_fullStr Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
title_full_unstemmed Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
title_short Impact of 3D cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on CAR-T cell activation
title_sort impact of 3d cell culture hydrogels derived from basement membrane extracts or nanofibrillar cellulose on car t cell activation
topic Biological sciences
Immune response
Cell biology
Materials science
Biomaterials
url http://www.sciencedirect.com/science/article/pii/S2589004225014956
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