人表皮黑素细胞与角质形成细胞共培养体外模型中黑素小体转运的观察

Establishment of the in vitro model of the melanocyte and keratinocyte co-culture system and observation of melanosome transfer

目的:建立人表皮黑素细胞与角质形成细胞共培养的体外模型,并且在模型中观察黑素小体的转运。方法:分别培养人表皮黑素细胞和角质形成细胞,用二乙酰羧基荧光素-琥珀酰亚胺酯(CFDA-SE)标记黑素细胞内黑素小体,然后将两种细胞共培养,并在倒置显微镜和共聚焦显微镜下观察共培养模型中黑素小体由黑素细胞向角质形成细胞的转运。结果:培养至第3天和第7天可在倒置显微镜下观察到黑素细胞与角质形成细胞共同存活,并通过树突发生联系,在共聚焦显微镜下可直接观察到黑素小体由黑素细胞向角质形成细胞的转运。结论:成功建立黑素细胞与角质形成细胞共培养的体外模型,并利用CFDA-SE标记,在共聚焦显微镜下直接观察到黑素向角质形成细胞的转运。

Objective： To establish an in vitro model of the melanocyte and keratinocyte co--culture system, then the melanosome transfer was observed in this model. Methods： The human epidermal melanocyte and keratinocyte were first cultured separately. Then the melanosomes in the melanocytes were fluorescence labeled with carboxy fluorescein diacetate ester （CFDA--SE）, keratinocytes and the labeled melanocytes were co-cultured for up to 7 days, and morphological manifestation of melanosomes transferred from keratinocytes to melanocytes was observed by both inverted and confocal microscopy. Results： On the 3rd day and the 7th day, live melanocytes and the keratinocytes were observed in the co-culture system and dendritic contact was observed between them by inverted microscopy. Moreover, the transfer of melanosomes from the melanocytes to the keratinocytes was observed by dint of fluorescence CFDA-SE under confocal microscope. Conclusions： The in vitro model of the melanocyte and keratinocyte co-culture system was successfully established and the transfer of melanosomes from melanocytes to keratinocytes is observed by using CFDA-SE staining.