家兔毛乳头细胞的分离、培养和鉴定  

Isolation, Culture and Identification of Rabbit Dermal Papilla Cells

作　　者：李佳丽[1] 刘明 周娟[1] 陈秋燃 陈阳[1] 陈实 申金钰 吴信生[1] Li Jia-li;Liu Ming;Zhou Juan;Chen Qiu-ran;Chen Yang;Chen Shi;Shen Jin-yu;Wu Xin-sheng(College of Animal Science and Technology Yangzhou University,Yangzhou 225009)

机构地区：[1]扬州大学动物科学与技术学院,扬州225009

出　　处：《中国养兔》2020年第3期4-7,16,共5页Chinese Journal of Rabbit Farming

基　　金：现代农业产业技术体系建设专项资金项目(CARS-43-A-1)。

摘　　要：毛囊经历周期性生长期,受其基部的真皮乳头细胞调节。本研究用显微切割和二步酶消化相结合的方法处理家兔触须,分离、培养毛乳头细胞,并通过碱性磷酸酶活性检测和蛋白免疫印迹对分离的细胞进行鉴定。结果发现,分离的原代毛乳头细胞呈现向四周放射性贴壁生长,且以长梭形纤维样多极向排列;低代数的毛乳头细胞有明显的凝集性现象,随着代数的增加,凝集性现象越来越不显著,至第10代凝集现象消失,第15代左右细胞停止增殖;经检测,分离的毛乳头细胞有很高的碱性磷酸酶活性,能够表达毛乳头细胞标志性蛋白α-SMA和VIM,表明所分离细胞为家兔的毛乳头细胞,为后期开展毛囊发育的生物学研究提供体外模型。Hair follicles undergo cyclical periods of growth, which are regulated by dermal papilla cells at the base ofthe follicle. In this study, the rabbit vibrissae was treated by microdissection and two-step enzymatic digestion. Thedermal papilla cells were isolated and cultured, and these cells were identified by alkaline phosphatase activity andwestern blot. The results showed that the isolated primary dermal papilla cells showed a radioactive adherent growthto the surrounding area and arranged in a multi-polar orientation of the long spindle-shaped fiber. The low-passagedermal papilla cells showed obvious aggregative behavior. With the increase of passage, the aggregative behavior be-came less and less significant. Until the passage 10, the aggregative behavior was complete invisible, and the dermalpapilla cells ceased proliferation around passage 15. The results showed that the isolated dermal papilla cells hadhigh alkaline phosphatase activity and could express the marker proteins alpha-SMA and VIM, which indicated thatthe isolated cells were dermal papilla cells of rabbits and provided an in vitro model for the biological research ofhair follicle development.

关 键 词：兔 毛囊 毛乳头细胞 二步酶消化法 

分 类 号：S829.1[农业科学—畜牧学]