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目的:建立最佳的口腔扁平苔藓(OLP)角质形成细胞的体外培养方法和条件,探索抗真菌治疗、年龄因素对其培养成功率的影响。方法:将网纹型OLP患者按照是否进行抗真菌治疗和年龄分组,胰酶消化法获取原代角质形成细胞(经病理确诊)。口腔黏膜上皮细胞培养基(OKM)进行体外原代及传代培养。Fisher确切概率法比较不同情况下的细胞培养成功率,倒置相差显微镜下观察细胞形态学变化。结果:抗真菌治疗后、年龄≤40岁者,其角质形成细胞的培养成功率明显高于未使用抗真菌措施及年龄>40岁者,差别具有统计学意义(P*=0.03,P**=0.015)。结论:OLP患者经过抗真菌治疗后以及选择年龄较轻的口腔黏膜组织在体外培养的成功率更高,进一步完善了人OLP角质形成细胞系的体外建立方法。
OBJECTIVE: To establish the best method and conditions for culturing the oral lichen planus (OLP) keratinocytes in vitro and to explore the effect of antifungal therapy and age on the success rate of the culture. Methods: The primary keratinocytes were obtained from the patients with veins of OLP according to their antifungal therapy and age groups and pancreatic enzyme digestion (confirmed by pathology). Oral mucosal epithelial cell culture medium (OKM) for primary and subculture in vitro. Fisher exact probability method to compare the success rate of cell culture under different conditions, inverted phase contrast microscope to observe the changes in cell morphology. Results: After antifungal therapy, the success rate of keratinocytes cultured in patients ≤40 years of age was significantly higher than those without antifungal and age> 40 years (P * = 0.03, P ** = 0.015). CONCLUSIONS: The success rate of OLP patients after anti-fungal treatment and the selection of younger oral mucosa cultured in vitro is further improved, further improving the establishment method of human OLP keratinocyte line in vitro.