摘要
目的建立毛囊隆突处外根鞘细胞的分离和培养方法。方法利用中性蛋白酶分离得到单个毛囊,采用胰酶消化法进行细胞培养,倒置6显微镜观察细胞形态,绘制细胞生长增殖曲线,免疫荧光检测干细胞相对特异性标识分子角蛋白19(K19)和β1整合素表达情况,分别用分离培养的角质形成细胞(Keratinocytes,KCs)做对照。结果免疫荧光检测显示90%左右的毛囊外根鞘细胞胞浆中均有K19和β1整合素的阳性表达。结论应用中性蛋白酶和胰酶消化法体外培养毛囊隆突处外根鞘细胞简单、可行。
Objective To establish a method for isolation and cultivation in vitro of human hair follicle bulge outer root sheath cells. Methods The single hair follicle were obtained by incubating with dispase, then harvest the outer root sheath cells from enzymic digestion and explant culture. Cells were observed by phase contrast microscope, meanwhile growth curve was made, immunofluoreseent assay to detect the expression of stem cell specific marker-cytokeratin 19 （CK19） and integrin 131. Results Human hair bulge follicle outer root sheath cells can be obtained from enzymic digestion and explant culture. Conclusion Enzymic digestion method is feasible in cultivation of human hair follicle outer root sheath cells in vitro.
出处
《中国现代医生》
2013年第6期1-3,6,F0003,共5页
China Modern Doctor
基金
国家自然科学基金(81071294)
浙江省自然科学基金(Z2100973)
浙江省杭州市科委医学重点专科专病专项资金(20091233Q09)
关键词
毛囊
外根鞘细胞
毛囊干细胞
角质形成细胞
分离
培养
Hair follicle
Outer root sheath cells
Hair follicle stem cell
Keratinocyte
Isolation
Cultivation
