人表皮黑素细胞与HaCaT细胞共培养体外模型的建立

Establishment of the co-culture model of human melanocyte and HaCaT cell in vitro

目的:建立人表皮黑素细胞与HaCaT细胞共培养的体外模型,观察α-促黑素以及烟酰胺对黑素在两细胞间传递的影响。方法:分别培养人表皮黑素细胞和HaCaT细胞,接种黑素细胞48h后再将HaCaT细胞以1:2的比例与黑素细胞共培养,分别以不同浓度的α-促黑素和烟酰胺干预共培养的细胞9天,Fontana-Masson银染法观察共培养模型中黑素颗粒在两细胞间传递的情况。结果:培养3天即可观察到约20%HaCaT细胞核周围出现从邻近黑素细胞传递来的黑素颗粒,随着时间的延长阳性染色HaCaT细胞比例逐渐增多,第7天达到高峰(约80%),α-促黑素以及烟酰胺分别以浓度依赖方式促进/抑制了黑素颗粒在两细胞间的传递。结论:成功建立了人表皮黑素细胞与HaCaT细胞共培养体外模型,为大规模筛选促/抑黑素传递的药物奠定了实验基础。

Objective To establish an in vitro model of melanocyte and HaCaT cell co-culture model, and measured the melanin transfer effects induced by α-MSH and niacinamide using the model. Methods Human epidermal metanocyte was co-culture with HaCaT cell as 1：2 ratio after 48 h of melanocyte first seeding on the dishes. Then melanin transfer was observed with Fontana-Masson method bya-MSH and niacinamide after 1,3,5,7,9 day of co-culture separately . Results About 20% HaCaT cells that have positive melanin stain around nuclear were observed after 3 day＇s co- culture. With the time developed, the ratio of positive melanin stained HaCaT cell is increasing and reached its peak （80%） at the 7th day. In our model, the dose-dependent inhibitory and facilitative effects were observed respectively in niacinamide and α-MSH treated group. Conclusions The in vitro model of epidermal melanocyte and HaCaT cell co- culture system was successfully established and it offers a new, suitable, reliable and easy tool for the clinical evaluation of melanin transfer modulators.