Figure 3.

Organoid cyclic stretch model. A: a 3-D-printed base has four cut nylon sponges placed in the four arms of the insert cross. B: a 3-D-printed lid has 8 radial spokes. C: after filling the mold with high-grade silicone and curing overnight, the insert is ready to be sterilized in 70% ethanol for use. D: after placing on a sterilized piece of parafilm, 50–100 µL of Matrigel is added. E: using a cut pipette tip, a single mouse lung organoid is placed in the well of the insert with minimal PBS and then covered with an additional 50–100 µL of Matrigel from the spaces between the spokes to keep the organoid in the well of the insert. F: the insert is placed into the biaxial stretching mount of a CellScale A2. G: schematic showing the dimensions of the silicone mold. H: 3-D-image of a membrane-stained mLO before stretch. Scale bar = 250 µm. Organoid dimensions are shown. (I) Image of the same organoid with static hold of 10.25% biaxial stretch. Scale bar = 250 µm. Quantification of the absolute (J) and relative change (K) in mLO volume with biaxial stretch. mLO, mouse lung organoid; 3-D, three-dimensional.