大鼠颌下腺上皮细胞原代培养转染实验  

Establishment of Primary Cultures of Rat Submandibular Gland Epithelial Cells for Transfection Expriment

作　　者：汤晓飞[1] 高文涛[1] 孙宏晨[1] 张泽兵[1] 徐辉[1] 欧阳喈[1] 

机构地区：[1]吉林大学口腔医学院病理科,吉林长春130041

出　　处：《口腔医学研究》2004年第2期149-152,共4页Journal of Oral Science Research

摘　　要：目的 :转染质粒pSV2neo、pSV3neo ,使大鼠颌下腺上皮细胞永生化 ,进而建立大鼠颌下腺肌上皮细胞系。方法 :采用改良磷酸钙沉淀法 ,将制备的质粒pSV2neo、pSV3neo转染大鼠颌下腺上皮细胞 ,G4 18筛选 ,Southern印迹杂交、透射电镜、免疫组化及软琼脂培养检测所筛选阳性细胞的性质。结果 :经质粒pSV2neo、pSV3neo转染分离的大鼠颌下腺上皮细胞仍保留上皮细胞的形态 ,生存期延长。经pSV3neo转染的上皮细胞 ,Southern印迹杂交证明整合有SV4 0T抗原 (LT) ;免疫组化和透射电镜观察显示存在有大量的肌上皮细胞和少量的腺泡细胞 ;软琼脂培养无克隆形成。结论Objective: To transfect rat submandibular gland epithelial cells with plasmid vectors and develop immortalized epithelial cells. Methods: The epithelial cells were transfected with plasmids pSV2neo、pSV3neo using modified CaPO4 precipitation. Results: The clonal epithelial cells retained epithelial cell differentiation. Southern blot hybridization showed that SV40 T gene was integrated into the genome of the epithelial cells transfected by pSV3neo, Ultrastructure and immunohistochemistry identified myoepithelial and acinar cells in the cells. There was no cell clone observed in soft agar. Conclusions: The clonal epithelial cells transfected by pSV3 neo were of immortalized inclination.

关 键 词：大鼠 颌下腺 肌上皮细胞 原代培养 胶原酶消化法 透射电镜 免疫组织化学 

分 类 号：R329-33[医药卫生—人体解剖和组织胚胎学]