Extended Data Fig. 10 | Perspectives for mimicking organ-level complexity
in mini-gut tubes through spatially controlled co-cultures. a, A schematic
representation of the 3D hydrogel-containing mini-gut chip (left). A cross-
sectional view (right) highlights the solid membrane-free co-culture, in a
biomimetic 3D environment, of an epithelium and various non-parenchymal
cell types seeded in the matrix surrounding the epithelium. b, Development of
engineered blood vessel-like networks composed of human endothelial cells.
Scale bars, 100 μm. c, Co-culture experiment of mini-gut tubes with
macrophages embedded into the surrounding hydrogel. Time-lapse imaging
revealed a direct interaction of highly motile macrophages with intestinal
epithelia in long-term co-cultures (here: 20 days). From day 2 onwards, the
macrophages acquire a distinct elongated morphology and migrate towards
the epithelium. Scale bars, 50 μm. d, Macrophages perform their phagocytic
function in a co-culture with mini-guts. Higher magnification views showing a
macrophage ingesting a particle from the basal side of the epithelium by
phagocytosis. Scale bars, 10 μm. e, Mini-gut tubes co-cultured with mouse
intestinal myofibroblasts incorporated in the surrounding matrix.
Myofibroblasts extensively migrate through the gel, extend processes and
directly interact with the epithelium. Scale bars, 50 μm. f, A co-culture
experiment of mini-guts epithelium with myofibroblasts initially co-seeded
together with ISCs directly into the lumen. Myofibroblasts rapidly attach
in the microchannel and remain incorporated into the continuous
monolayer generated by ISCs. From day 3 onwards, myofibroblasts, localized
predominantly in crypt regions, extended pseudopodia into the surrounding
matrix (arrows), recapitulating the in vivo tissue architecture. Scale bars,
50 μm. All data are representative of at least two independent experiments.
EDF of bright-field images (c–f), calculated for a z-stack of 80 μm.