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. 2018 Aug 23;3(16):e122211. doi: 10.1172/jci.insight.122211

Figure 4. IPF cultured epithelial cell–conditioned supernatants promote lung fibroblast invasion.

Figure 4

Normal and IPF explant suspensions were cultured in PEB medium until epithelial cell colonies were apparent. After 2–3 passages, cells were cultured in 6-well plates for 24 hours, after which culture supernatants (CSs) were collected and frozen at –20°C. Lung fibroblasts were then plated into 96-well plates, scratched using a WoundMaker, and treated with normal or IPF epithelial cell CS mixed with Matrigel. Lung fibroblast invasion was monitored using an Incucyte Zoom live cell imager. (A and B) Depicted are representative images showing the closure of wounded areas (blue) by normal (n = 1; A) or IPF (n = 2; B) lung fibroblasts (yellow) after treatment with normal (n = 2; top) or IPF (n = 4; bottom) epithelial cell CS over the span of 48 hours. (C–E) Depicted is the average wound closure (relative to the initial wound) over 90 hours of normal (n = 1) or IPF (n = 2) fibroblasts treated with 2 normal (red) or 4 IPF (black) PEC CSs. Data shown are the mean ± SEM. PEB, PneumaCult-Ex Plus Medium + 10 µM Y27632; PEC, pulmonary epithelial cells; IPF, idiopathic pulmonary fibrosis.