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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 5 μg/mlHC的DMEM培养基最适合体外原代及传代培养人胚颌下腺细胞
OBJECTIVE: To study the methods of primary and subculture of human submandibular gland cells and lay a foundation for further study on the function, origin, occurrence and development of submandibular gland cells. Methods: FBS, epidermal growth factor (EGF) and hydrocortisone (HC) were added into DMEM medium to observe the growth characteristics of the cells. RESULTS: Epithelial cells grew best in DMEM containing φ = 2% FBS, 10 ng / ml EGF, 5 μg / ml insulin (INS) and 5 μg / ml HC and poorly growing epithelial cells in the other groups. The cultured cells are round, triangular, polygonal and have good cell extension. The cells grew well after passage, the morphology did not change. HE staining shows cells are round, triangular, polygonal, nuclear blue dye, cytoplasm pink dye. Most cytokeratin staining positive cells, described as epithelial cells, a small number of cells CK18 staining positive, indicating that cultured cells contain a portion of ductal epithelial cells. CONCLUSION: DMEM containing φ = 2% FBS, 10 ng / ml EGF, 5 μg / ml INS and 5 μg / ml HC is the most suitable primary and subcultured human embryonic submandibular gland cells