Journal: Nature Biomedical Engineering

Article Title: Bioengineered immunocompetent preclinical trial-on-chip tool enables screening of CAR T cell therapy for leukaemia

doi: 10.1038/s41551-025-01428-2

Figure Lengend Snippet: a , The compartmentalized bone marrow chip (middle) was populated with human bone marrow cells (right) to replicate the in vivo counterpart (left). The whole scanning of the leukaemia bone marrow chip (bottom right) with central sinus (RFP-HUVECs in red), medullary cavity and endosteum (DiD-labelled osteoblasts in yellow). The 3D view of perivascular niche (top right) in bone marrow with hematopoietic cells (CD45 + in red), vascular cells (CD31 + in orange) and nuclei in blue (DAPI). Representative images were from one of the three technical replicates with similar results ( n = 3). Schematic (left part) was adapted from smart.servier.com . Schematic (right part) was created in BioRender (C.M. (2025), https://BioRender.com/wlbhoxu ), b , scRNA-seq profiling of the bone marrow cellularity on-chip, highlighting the presence of most hematopoietic (immune) and non-hematopoietic (bone marrow stroma) cells. c , scRNA-seq profiling of primary bone marrow mononuclear cells, which was comparatively mapped to that of on-chip bone marrow niche. d , The cellularity where B cell populations were excluded to reveal bone marrow niche immune populations. In addition to the bone marrow stromal cell populations seeded to build the stromal environment, macrophage, basophil/mast cells and megakaryocyte/platelets were generated on-chip during culture. e , The presence of stromal compartment. HUVECs in red (RFP), Reh B-ALL cells in green (GFP) and mesenchymal cells in purple (DiD labelling). The white arrowheads indicate Reh B-ALL cells. f , The presence of hematopoietic cells. Lymphoid (left): CD8 + T cells in cyan and CD4 + T cells in red. The top white arrowheads indicate CD4 + T cells, and the bottom arrowheads indicates CD8 + T cells. Myeloid (right): monocyte (CD14 + ) in cyan, HUVECs (CD31 + ) in green and Reh B-ALL cells (CD19 + ) in red. The white arrowheads indicate monocytes. g , The deposition of ECMs such as laminin (green) and collagen IV (purple). Reh B-ALL cells in red. The white arrowheads indicate Reh B-ALL cells. Representative images were from one of the three technical replicates with similar results ( n = 3).

Article Snippet: Primary human umbilical vein endothelial cells (HUVECs; catalogue number C2519A, Lonza), VE-CAD-GFP-expressing HUVECs (catalogue number cAP-0001VECAD-GFP, Angio-Proteomie) and RFP-expressing HUVECs (catalogue number cAP-0001RFP, Angio-Proteomie) were cultured in Endothelial Cell Growth Medium-2 BulletKit (EGM-2; catalogue number CC-3162, Lonza) and used within passage 5.

Techniques: In Vivo, Generated