Fig. 6: Organoids’ growth and maturation enhancement through flow and vascularization.

a, b BVO (brightfield, dotted cyan line) and HUVEC network (GFP) development from day 0 to day 10 on-chip, in static (a) and flow (b) conditions (diagrams created with BioRender.com). Enhanced ECM remodeling under flow conditions can be observed in the brightfield images at day 10, particularly by the presence of black structures surrounding the trap site, indicative of active cell processes (highlighted by white arrows). c Angiogenesis Analyzer outputs of the morphology of the endothelial networks between static and flow conditions after 10 days of culture on-chip (n = 8 (static) and 10 (flow)). d BVOs’ growth after 10 days of culture on-chip in various conditions, reported as a percentage increase in their size (average of minor and major axes from the fitted ellipse) compared to day 0 (n = 7 (static w/o vasc.), 11 (static with vasc.), 7 (flow w/o vasc.) and 12 (flow with vasc.)). e Volcano plot showing differentially expressed genes between conditions static w/o vasc. and flow with vasc. f Most enriched (top 15) Gene Ontology (GO) biological process terms resulting from the comparison between conditions static w/o vasc. and flow with vasc. g Heatmap of a selection of genes associated with ECM organization (dotted green boxes) and blood vessel development (dotted black boxes) pathways. The color-coded representation illustrates the expression patterns of key genes involved in these biological processes, enabling a visual comparison of their relative expression levels between conditions static w/o vasc. and flow with vasc. h Classification of BVO transcriptomes with a reference tissue simulated from single-cell RNA-seq data of BVOs matured in mice. Semi-supervised classification based on the Kruskal-Wallis test of genes significantly differentially expressed across all conditions, followed by visualization of the top 50 most significant genes (diagrams created with BioRender.com). Scale bars, 200 µm (a, b and d). Data represents mean ± s.d. Statistical significance was attributed to values of P < 0.05 as determined by unpaired t test (two-tailed). *P < 0.05 (P = 0.05 (static w/ vasc. vs flow w/o vasc.) and P = 0.03 (static with vasc. vs. flow w/o vasc.)), **P < 0.01 (P = 0.008), ***P < 0.001. Source data are provided as a Source Data file.