人表皮黑素细胞及HaCaT细胞共培养体外模型建立.docVIP

人表皮黑素细胞及HaCaT细胞共培养体外模型建立[摘要]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。 [关键词]黑素细胞；共培养；HaCaT细胞；黑素传递 [中图分类号]Q813.1[文献标识码]A[文章编号]1008-6455（2009）02-0188-03 Establishment of the co-culture model of human melanocyte and HaCaT cell in vitro MA Hui-jun, TIAN Yan, LIU Wen, LI You, ZHAO Guang,YANG Qing-qi (Department of Dermatology, The Airforce General Hospital of PLA, Beijing 100142,China) Abstract:ObjectiveTo 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.MethodsHuman epidermal melanocyte 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 byα-MSH and niacinamide after 1,3,5,7,9 day of co-culture separately . ResultsAbout 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.ConclusionsThe 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. Key words：melanocyte; co-culture; HaCaT cell; melanosome transfer 表皮中的黑素细胞和角质形成细胞相互作用共同组成了表皮黑素单元，起着重要的调节皮肤颜色、防止紫外线辐射的作用。目前研究表明，人皮肤颜色主要受色素沉着过程的影响，除黑素细胞生成黑素外，黑素小体从黑素细胞向周围角质形成细胞的广泛传递过程在皮肤颜色调节中起着更为重要的作用[1]。目前黑素传递的研究