正常人角质形成细胞和黑素细胞体外共培养体系的建立  

Establishment of coculture system of normal human epidermal keratinocytes and melanocytes in vitro

作　　者：李海东[1] 王鹰[2] 

机构地区：[1]中国人民解放军第三二四医院呼吸科,重庆400020 [2]中国人民解放军第三二四医院皮肤科,重庆400020

出　　处：《重庆医学》2012年第8期769-771,F0003,F0002,共5页Chongqing medicine

基　　金：重庆市自然科学基金资助项目(CSTC2009BB5313)

摘　　要：目的体外建立正常人表皮角质形成细胞和黑素细胞直接接触的共培养体系。方法分别培养角质形成细胞和黑素细胞,体外建立角质形成细胞和黑素细胞直接接触的共培养模型,采用四甲基偶氮唑盐(MTT)比色法、紫外分光光度计检测、左旋多巴(L-Dopa)染色、免疫组织化学染色及透射电镜观察对共培养体系中的黑素细胞和角质形成细胞进行生物学鉴定。结果单独培养时,角质形成细胞呈圆形、"铺路石样"生长,黑素细胞呈两极或多级树突状生长;角质形成细胞和黑素细胞按一定比例混合培养后,2种细胞均迅速增殖,黑素细胞树突多为3～5个,与数十个角质形成细胞呈团块状生长,形成类似黑素单元的结构。共培养体系中的黑素细胞和角质形成细胞具备正常的生物学功能。结论角质形成细胞和黑素细胞直接接触的共培养体系为黑素细胞提供了更接近生理状态的体外研究模型。Objective To establish coeulture system of direct cell-to-cell contact between normal human epidermal keratinoeytes and melanocytes in vitro. Methods Keratinocytes and melanocytes were cultured respectively,and then the coculture model of direct cell-to-cell contact between keratinocytes and melanocytes was established in vitro, methyl thiazolyl tetrazolium（MTT） assay, UV speetrophotometry,levoDopa（L-Dopa） staining, immunohistochemieal staining and transmission electron microscopy observation were employed to biologically identify the melanoeytes and keratinoeytes in the coculture system. Results Keratinocytes were round and grew in ＂cobblestone＂ pattern while melanoeytes in bipolar or multi-level dendritic growth pattern when they were cultured separately. The 2 kinds of cells proliferated rapidly when they eocuhured in a certain proportion. Melanocytes with 3 to 5 dendrites grew in crumb pattern with dozens of keratinoeyte,similar to the structure of melanin unit. Melanocytes and keratinocytes in the coculture system exhibited normal biological functions. Conclusion Coeulture system of direct cell to cell contact between normal human epidermal keratinocytes and melanoeytes provides a model closer to physiological condition for melanocytes study in vitro.

关 键 词：黑素细胞 角质形成细胞 共同培养技术 生物学鉴定法 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]