人口腔扁平苔藓角质形成细胞的体外培养及生物学鉴定

Culture and biological identification of human oral lichen planus keratinocytes in vitro

目的建立人口腔扁平苔藓（orallichenplanus，OLP）角质形成细胞体外培养方法，观察其生物学特性。方法选择组织病理学确诊的OLP患者黏膜组织，无血清角化细胞培养基进行体外原代及传代培养，倒置相差显微镜下观察细胞形态学变化，角蛋白间接免疫荧光鉴定和电镜观察细胞超微结构。结果成功培养了Ⅰ型OLP（网状、斑块型）角质形成细胞，连续传代5～6代，整个细胞生长期间为单一的角质形成细胞，呈典型的铺路石状。角蛋白间接免疫荧光反应阳性，透射电镜显示：细胞胞浆内可见大量的空泡性结构，与OLP的病理学特征一致，能够鉴定我们培养的细胞为OLP角质形成细胞。结论本研究成功建立了体外培养人OLP角质形成细胞的方法，为人OLP细胞模型的建立打下坚实基础。

Purpose To establish a method for culturing human oral lichen planus（OLP） keratinocytes in vitro and observe its biological characteristics. Methods Specimens obtained from OLP patients by pathology final diagnosed were cultured in serum-free keratinocyte medium. Morphological characteristics were observed under inverted phase contracted microscope. The cultured cells were i- dentified by immunofluorescence staining with monoclonal cytokeratin antibody. Uhrastructure of cell was observed by electron microscopy. Results We have cuhulred type Ⅰ OLP keratinocytes successfully in vitro. The cells could be maintained in culture up to 5 - 6 passages. Immunofluorescence showed that the cultured cells were positive to cytokeratin antibody. Transmission electron microscope manifested that there was many vacuolus in the intracytoplasm of OLP keratinocytes, which conformed to pathology characteristic of OLP, and we can identify the cells were OLP keratinocytes. Conclusions The method used in this work for cultivation of type Ⅰ OLP keratinocytes is successful. It is an important base for construction of OLP cell model.