The Culture and Identification of Melanocytes in Melasma Skin

Objective To investigate the culture method of melanocytes in melasma skin, and observe the morphology and biological properties of the cultured melanocytes in vitro. Methods The skin specimen was obtained from melasma patients after lower blepharoplasty. Epidermis was isolated by 0.25% dispase, and then single cell suspension was obtained after digested by 0.25% trypsin and 0.02%EDTA. M254/HMGS conditioned media was added to cultured the cells purifiedly, the cell morphology was observed through inverted microscope, S-100 protein immunohisto chemical staining and dopa-staining was used for identification. The TRP-1, HMB-45 expression were studied by indirect immunofluorescence. Results The human melanocytes in melasma skin were successfully cultured, and the cells were multi-dendritic. S-100 protein immunohisto chemical staining and Dopa staining were positive. The TRP-1 and HMB-45 were positive expression by indirect immunofluorescence. Conclusion Melanocytes can be successfully cultured in melasma skin with conditioned media insisted of special ingredients, which can be used for further investigation.