鼓膜上皮细胞和成纤维细胞的培养与鉴定

Isolation of pure tympanic membrane epithelial cells and fibroblasts

目的:探讨如何以简便快速的方法获得可以用于组织工程鼓膜构建的高纯度的鼓膜上皮细胞和成纤维细胞.方法: 分离新生豚鼠的鼓膜组织,利用本实验建立的两步组织块培养法获得纯化的鼓膜上皮细胞和成纤维细胞,倒置显微镜观察细胞生长状态,免疫组织化学染色鉴定细胞来源.结果: 用该方法可以获得纯化的鼓膜上皮细胞和成纤维细胞,细胞产量大,纯度高,鼓膜上皮细胞细胞具有典型的铺路石样形态,成纤维细胞呈梭形生长. 结论: 本实验所建立的两步组织块培养法可以获得大量高纯度的鼓膜上皮细胞和成纤维细胞以用于组织工程鼓膜的构建.

AIM: To develop a rapid method for the isolation of pure epithelial cells and fibroblasts from tympanic membrane. METHODS: The epithelial cells and fibroblasts were isolated and purified from tympanic membrane by a new two-step method used in our lab. The cells were identified by cell morphorlogy and immunostaining. RESULTS: Our cultured tympanic membrane epithelial cells were of characteristic cobblestone morphology and showed a positive reaction keratin. The fibroblasts cultured were spindle-shaped and showed a positive reaction to antibodies against vimentin. The tympanic membrane epithelial cells and fibroblasts cultures of this processing were free of any contaminants. CONCLUSION: Pure tympanicmembrane epithelial cells and fibroblasts can be easily isolated by our method.