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The serum-free culture supernatant derived from EBC-1 cells (EBC-1 supernatant) and the cells were spun down, and each supernatant was filtrated through a 0.45-µm polyvinylidene difluoride membrane filter (Merck Millipore, Italy) and concentrated by ultrafiltration (Amicon Ultra 3K, Merck Millipore)