Figure 7

Wound healing and proliferation assay for HCECs depending on CM concentrations (× 0.5, × 1.0, × 3.0, × 5.0). Culture media for the HCEC migration assay was composed of HCEC medium containing supplements with various concentrations of CM (× 0.5, × 1.0, × 3.0, × 5.0) derived from the ADSC culture. After the transplanted cells reached confluence, the plates were scratched using a pipette tip and scratch gaps compared between the 0 h and 48 h time points (A,B). The equation for the percentage of migration was (100-wound area at 48 h/wound area at 0 h) × 100. To measure the proliferative capacity of the cells, the degree of cell viability was measured using an MTT [3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazoliumbromide] assay at 540 nm (C,D). CM × 0.5 more potently induced proliferation of HCECs over the other groups. After the migration and proliferation assay, HCECs were harvested and mRNAs for migration-related genes and epithelial growth cytokines were analyzed using PCR (E). (B) and (D), 100X magnification; MMP, matrix metallopeptidase; IL-6, interleukin-6; EGF, epithelial growth factor; HGF, hepatocyte growth factor; CDK, cyclin-dependent kinase; CXCR4, C-X-C chemokine receptor type 4; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.