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We present a protocol to generate immunocompetent 3D tumor-on-chip models from human solid tumors, enabling more accurate therapy response assessment than traditional 2D assays. We outline the isolation and culture of autologous tumor cells, CD8 tumor-infiltrating lymphocytes, and cancer-associated fibroblasts, followed by their encapsulation in a 3D biomimetic matrix within microfluidic devices and subsequent video microscopy. The protocol is adaptable to other tumor types, including breast and colon cancer. For complete details on the use and execution of this protocol, please refer to Veith et al..
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http://dx.doi.org/10.1016/j.xpro.2025.103895 | DOI Listing |
STAR Protoc
June 2025
Institut Curie, INSERM U1339, Stress and Cancer Laboratory, PSL Research University, Paris, France. Electronic address:
We present a protocol to generate immunocompetent 3D tumor-on-chip models from human solid tumors, enabling more accurate therapy response assessment than traditional 2D assays. We outline the isolation and culture of autologous tumor cells, CD8 tumor-infiltrating lymphocytes, and cancer-associated fibroblasts, followed by their encapsulation in a 3D biomimetic matrix within microfluidic devices and subsequent video microscopy. The protocol is adaptable to other tumor types, including breast and colon cancer.
View Article and Find Full Text PDFAdv Healthc Mater
May 2025
Institut Curie, U1339 INSERM - UMR3666 CNRS, Stress and Cancer Laboratory, PSL Research University, Paris, 75005, France.
The endothelium compartment is a key player in tumor initiation and progression, but most existing tumor-on-chip models lack clinical relevance. Here, a 3D vascularized tumor-on-chip (vToC) model, generated with patient-derived microvascular endothelial cells (ECs) that are freshly isolated from surgical lung cancer samples, is presented. The microvessel molecular identity, morphology, and functionality are assessed by transcriptomic, immunofluorescence, TNF-α stimulation, and permeability assays.
View Article and Find Full Text PDFAdv Drug Deliv Rev
June 2021
Department of Biomedical Science, Faculty of Medicine, Eberhard Karls University Tübingen, Österbergstraße 3, 72074 Tübingen, Germany; Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Nobelstrasse 12, 70569 Stuttgart, Germany. Electronic address: peter.loskill@uni-tuebinge
The advances in cancer immunotherapy come with several obstacles, limiting its widespread use and benefits so far only to a small subset of patients. One of the underlying challenges remains to be the lack of representative nonclinical models that translate to human immunity and are able to predict clinical efficacy and safety outcomes. In recent years, immunocompetent Cancer-on-Chip models emerge as an alternative human-based platform that enables the integration and manipulation of complex tumor microenvironment.
View Article and Find Full Text PDFCell Rep
December 2018
Institut Curie, Centre de Recherche, Paris Sciences et Lettres Research University, 75005 Paris, France; ART Group, INSERM U830, 75005 Paris, France. Electronic address:
A major challenge in cancer research is the complexity of the tumor microenvironment, which includes the host immunological setting. Inspired by the emerging technology of organ-on-chip, we achieved 3D co-cultures in microfluidic devices (integrating four cell populations: cancer, immune, endothelial, and fibroblasts) to reconstitute ex vivo a human tumor ecosystem (HER2 breast cancer). We visualized and quantified the complex dynamics of this tumor-on-chip, in the absence or in the presence of the drug trastuzumab (Herceptin), a targeted antibody therapy directed against the HER2 receptor.
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