Objective   To investigate the role of PKC-ζ activation in stromal-derived factor 1 (SDF-1) mediated migration of human epidermal stem cells and its underlying mechanism.    Methods   Epidermal stem cells were isolated from preputial skin samples by adhering to collagen type Ⅳ after digesting with Dispase Ⅱ and then cultured in vitro in serum-free defined keratinocyte medium (DK-SFM). Transwell migration assay was used to observe the migration of human epidermal stem cells that were treated with different PKC inhibitors

including chelerythrine chloride

staurosporine and PS-ζ. Western blot analysis was used to analyze the phosphorylation of PKC-ζ upon SDF-1 stimulation at different time phase. Phalloidin-Rhodamine was used to illustrate morphology of cell cytoskeleton by staining of the F-actin. Laser scanning confocal microscopy was used to observe the polarization of human epidermal stem cells following SDF-1 stimulation.    Results   Pretreatment of human epidermal stem cells with chelerythrine chloride or PS-ζ decreased the migration of human epidermal stem cells to SDF-1. However

treatment with staurosporine did not affect SDF-1-induced chemotaxis. SDF-1 induced PKC-ζ activation

which was demonstrated by the increased phosphorylation of PKC-ζ with time elapsing. PS-ζ inhibited the polarization of human epidermal stem cells induced by SDF-1.    Conclusion   SDF-1 promotes the directional migration of human epidermal stem cells via inducing the phosphorylation of signaling protein PKC-ζ and affecting the polarization of F-actin.