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RAW 264.7 cells were cultured in complete Dulbeccos modified Eagle medium (DMEM, Gibco) supplemented with 10% (v/v) heat-inactivated fetal bovine serum (FBS) (Avantor Seradigm), 2 mM L-glutamine (Gibco), 1 non-essential amino acids solution (Gibco) and 1 mM sodium pyruvate (Gibco) below 18 passages without exceeding 90% confluency

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Research demonstrates that GHK-Cu stimulation of collagen synthesis begins at concentrations between 10 and 10 M and maximizes at 10 M, with effects independent of changes in cell proliferation, indicating direct activation of synthetic machinery rather than simply increasing fibroblast numbers

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