An endothelial cell migration assay was performed by using Transwell chambers (pore size of 8 μm, 6.5 mm in diameter; Millipore, Billerica, MA). In quadruplicate, IECs or AECs (1.2 × 104 cells/well) were seeded into the upper chambers in 120 μl of DMEM or M199 without FBS or other supplements, and 500 μl of SC-CM-DMEM or SC-CM-M199 (DMEM or M199 as a control) containing 0.5% bovine serum albumin (BSA; Beyotime) and 1% FBS was added to the lower chambers. The upper chambers were removed from the lower chambers after incubation at 37 °C for 6 h and wiped with cotton swabs. The polycarbonate membranes were fixed using methanol and stained with crystal violet. The membranes were eluted by using 100 μl of 10% acetic acid. The optical density (OD) of the eluted fluid in triplicate wells was measured at an absorbance of 570 nm.

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