大鼠舌背上皮细胞体外培养及其生物学特性观察

目的:探讨舌背上皮细胞体外培养的可行性,并观察其生长特性。方法:断颈处死出生后1d的SD大鼠,70%乙醇浸泡3～5min,切取舌体及下颌骨。以DMEM/F12作为基础培养液,原代培养舌背黏膜上皮。当细胞铺满整个培养瓶底70%时,进行细胞传代。采用广谱角蛋白细胞免疫化学染色法及透射电镜观察法对上皮细胞进行鉴定。结果:接种时,原代细胞形状、大小不等,呈多样性,表现为多角状扁平细胞、体积较大的球形细胞和体积较小的球形细胞。大多数细胞在培养24h后贴壁;培养3d后,细胞增殖形成许多小而不规则的细胞克隆或集落,开始进入指数增殖阶段,形成细胞团块。随着时间推移,细胞团块周围结构疏松,间隙增大,细胞团块不断增大,融合、连接成片,排列成铺路石状,可传3～4代。结论:以DMEM/F12为基础培养液,可成功地培养出舌背上皮细胞。

Primary culture and histomorphological study of lingual dorsal epithelial cells in rat

PURPOSE: To investigate the feasibility of primary culture of lingual dorsal epithelium of SD rat in vitro,and to study the histomorphological characteristics of lingual dorsal epithelium. METHODS: Tongues of ten SD rats sacrificed by cervical dislocation under sterile condition were dissected at the age of 1day. The primary cultures of lingual dorsal epithelia were incubated in Dulbecco's modified Eagle's medium/F12.Cell passage was carried out when the cells of the primary cultures spread over 70% area of the bottom. The epithelial cells were identified by immunocytochemical stain with broad-spectrum cytokeratin(CK) and observed under transmission electron microscope. RESULTS: The primary cells were different in size and polymorphic such as cerioid flat cells,large and small globular cells. After cultured for 24 hours,most epithelial cells attached to the bottom of containers. At the third day,the epithelial cells proliferated and formed cluster. Finally,the mass of the epithelia was confluent into monolayer with cobblestone-like at day 7 to 10. And these cells could be passaged for 3 generations. CK was detected positively by immunocytochemistry. Furthermore,keratin filament and macula adhesions were observed under electron microscope. CONCLUSION: It is feasible to culture the lingual dorsal epithelium in DMEM/F12.