人颌下腺腺上皮细胞体外培养及生物学特性研究

A Study of Biological Characteristics with the Cultured Epithelial Cells in Vitro of Human Submandibular Glands

目的 建立人颌下腺腺上皮细胞的原代培养模型。方法 取口腔癌患者未受累及的颌下腺组织块 ,采用机械法和消化法 ,在 10 %血清浓度下 ,分别加入EGF、胰岛素、转铁蛋白等附加因子 ,37℃ ,5 %CO2 开放培养。选用苏木精 -伊红、PAS染色及S -P法免疫组化染色 ,扫描及透射电镜观察等方法研究其生物学性状。结果 通过特殊染色、免疫组化、电镜观察证实了体外培养细胞的腺上皮属性 ,细胞在 4d内可保持分化状态和分泌功能 ,可存活 1周以上。

Objective To establish a primary culture method of the human submandibular gland epithelial cells and test their biological charcateristics.Methods The fragments from adult human submandibular glands were dissected and sequentially digested. The isolated cells were cultured at 37℃ with 10% heat inactivated Bovine serum, epidermal growth factor, insulin, et al. Cell proliferation, morphology and ultrstructure were observed by phase contrast microscope, cytologic enzyme staining and electronic microscope. The cultured cells were identified by histochemical and immunohistochemical methods.Results Examinations of the cultured cells demonstrated characteristics of epithelial cells. They were able to survive for 7*!d, and maintained significant biosynthetic activity for 4*!d.Conclusion We have established a successful model of human submandibular primary culture in vitro.