Data-independent acquisition (DIA) based quantitative proteomic analysis of dermal fibroblast supernatant samples

We collected fibroblasts derived from the foreskin of two healthy patients and cultured the cells in 7.5% HPL + DMEM medium. After in vitro expansion to passage 3 (P3), once the cells reached 80–90% confluency, the medium was replaced with an equal volume of fresh DMEM. The conditioned supernatant produced over a 3-day period was collected and subjected to proteomic sequencing to determine the protein composition of the fibroblast-derived supernatant.