关键词: 黑线仓鼠, 黑色素细胞, 分离培养

Abstract: Objective　To establish a method for the isolation and identification of melanocytes from the skin of albino mutant chinese hamsters (AL) and wild-type hamsters (WT), and to analyze the functions of melanin synthesis and cell proliferation, in order to provide an ideal primary cell model for the study of albinism pathogenesis in this spontaneous albino mutant animal model.Methods　Hamster back skin tissues within 3 days of birth were removed, and melanocytes were isolated by trypsin digestion and cultured in MelM medium. Melanocytes were identified by immunofluorescence staining, L-Dopa staining, and transmission electron microscopy.The melanin content was determined by Masson-Fontana staining and colorimetric assay,the tyrosinase (TYR) activity was detected by L-Dopa catalytic assay and MTT assy, the cell proliferation was determined by cell cycle analysis and colony formation assy.Results Immunofluorescence of melanocyte-specific proteins TYR and Pmel17 showed that isolated AL and WT skin melanocytes all expressed. L-Dopa staining of AL melanocytes was significantly lighter than that of WT melanocytes, and there were fewer intracellular maturation-stage melanosomes. Melanin content and TYR activity of AL melanocytes were significantly lower than that of WT melanocytes (P<0.001); the proliferation rate of AL melanocytes was significantly faster than that of WT melanocytes (P<0.05), the proportion of S-phase cells was significantly higher (P<0.05), and the colony-forming ability was significantly enhanced (P<0.01).Conclusion　Trypsin digestion can obtain functional melanocytes from albino mutant chinese hamster skin, which have weakened melanin synthesis but enhanced proliferation ability.

Key words: Chinese hamster, melanocytes, isolation and culture