人胚颌下腺细胞原代及传代培养  

Culture of human fetus submandibular gland cells in vitro

作　　者：白忠诚[1] 司徒镇强[2] 刘斌[2] 李莉莉[3] 温德升[2] 李洁[2] 李焰[2] 

机构地区：[1]西安第四军医大学口腔医学院口腔颌面外科,710032 [2]第四军医大学口腔医学院口腔生物学教研室 [3]第四军医大学口腔医学院修复科

出　　处：《实用口腔医学杂志》2004年第3期276-279,共4页Journal of Practical Stomatology

基　　金：国家自然科学基金资助项目;课题编号 :30 1 71 0 1 8

摘　　要：目的 :研究人胚颌下腺细胞原代及传代培养的方法 ,为进一步研究颌下腺细胞的功能、起源、发生及发展奠定基础。方法 :在DMEM培养基中加入FBS、表皮生长因子 (EGF)及氢化可地松 (HC) ,观察细胞的生长特点。结果 :在含有 φ =2 %FBS、10ng/mlEGF、5 μg/ml胰岛素 (INS)及 5 μg/mlHC的DMEM培养基中上皮细胞生长最好 ,其它组上皮样细胞生长状态不佳。培养的细胞为圆形、三角形、多边形 ,细胞伸展良好。细胞传代后生长良好 ,形态并未发生改变。HE染色可见细胞为圆形 ,三角形 ,多边形 ,核蓝染 ,胞浆粉染。大多数细胞cytokeratin染色阳性 ,说明为上皮细胞 ,少数细胞CK18染色阳性 ,说明培养的细胞中含有一部分导管上皮细胞。结论 :含有 φ =2 %FBS、10ng/mlEGF、5 μg/mlINS及 5Objective: To investigate the method of culture and subculture of human submandibular gland epithelial cells.Methods: Human fetus submandibular gland cells were cultured by explant techmque in DMEM medium supplemented with φ=5% or φ=2% of fetus bovine serum(FBS), 10 ng/ml of epidermal growth factor(EGF),5 μg/ml of insulin (INS) and 5 μg/ml of hydxocotisone(HC). Subculture was conducted by trypsinization. Cells were identified by AE1/AE3 staining and CK18 staining. Results:Epithelial cells grew better with round, triangle and polygon morphology in DMEM medium supplemented with φ=2% FBS, 10 ng/ml EGF, 5 μg/ml INS and 5 μg/ml HC. Subcultured cells grew well and maintained epithelial. HE staining showed that the cells were round, polygon and triangle, nuclei were stained blue and cytoplasm were stained red. Immunohistochemical staining showed that most cultured cells were AE1/AE3 positive epitheliod cells, and there were CK18 positive cells (duct cells). Conclusion: DMEM medium supplemented with φ=2%FBS, 10 ng/ml EGF , 5 μg/ml INS and 5 μg/ml HC is feasible for the culture and subculture of human fetus submandibular gland cells.

关 键 词：胚胎 颌下腺 原代培养 传代培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]