Protocol for the biofabrication of immunocompetent tumor-on-chip models from patient solid tumors for assessment of anticancer therapies.

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.

Bioengineering a Patient-Derived Vascularized Lung Tumor-on-Chip Model to Decipher Immunomodulation by the Endothelium.

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.

Protocol for in vitro assessment of human monocyte transendothelial migration using a high-throughput live cell imaging system.

In vitro modeling of the different steps of immune cell recruitment is essential to decipher the role of endothelial cells in this process. Here, we present a protocol for the assessment of human monocyte transendothelial migration using a live cell imaging system. We describe steps for culture of fluorescent monocytic THP-1 cells and chemotaxis plate preparation with HUVEC monolayers.

A role for endothelial alpha-mannosidase MAN1C1 in radiation-induced immune cell recruitment.

Radiation therapy damages tumors and normal tissues, probably in part through the recruitment of immune cells. Endothelial high-mannose N-glycans are, in particular, involved in monocyte-endothelium interactions. Trimmed by the class I α-mannosidases, these structures are quite rare in normal conditions.

Deep models of integrated multiscale molecular data decipher the endothelial cell response to ionizing radiation.

The vascular endothelium is a hot spot in the response to radiation therapy for both tumors and normal tissues. To improve patient outcomes, interpretable systemic hypotheses are needed to help radiobiologists and radiation oncologists propose endothelial targets that could protect normal tissues from the adverse effects of radiation therapy and/or enhance its antitumor potential. To this end, we captured the kinetics of multi-omics layers-i.