大鼠颌下腺细胞培养方法建立及生物学特性研究  

Establishment of culture method of rat submandibular gland cells and their biocharacteristics

作　　者：刘超[1] 苗雷英[2] 孙宏晨[3] 乔春燕[3] 刘金钟[3] 柯小亮[3] 

机构地区：[1]吉林大学口腔医院正畸科,吉林长春130041 [2]吉林大学口腔医院牙体牙髓病科,吉林长春130041 [3]吉林大学口腔医院口腔病理学教研室,吉林长春130041

出　　处：《吉林大学学报（医学版）》2009年第2期218-221,F0002,共5页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(30672338)

摘　　要：目的:建立大鼠颌下腺细胞体外培养的方法,为涎腺细胞的体外扩增及基因治疗涎腺疾病奠定基础。方法:无菌条件下取1日龄Wistar大鼠颌下腺,经LB3、胰酶联合消化后原代细胞接种于含2%胎牛血清、表皮生长因子、胰岛素等的DMEM/F12培养液,相差显微镜观察原代及传代细胞的形态特征,免疫组织化学染色鉴定细胞来源,PAS染色观察体外培养第2代细胞合成、分泌多糖功能。结果:体外培养的大鼠颌下腺原代细胞为圆形、三角形、多边形。细胞传至第2代生长良好,形态未发生改变。体外培养的第2代细胞角蛋白、E-钙联蛋白免疫组织化学染色、PAS染色均为阳性,表明细胞为上皮来源且具有合成、分泌多糖类物质的功能。结论:酶联合消化法成功地分离、体外培养了大鼠颌下腺细胞,第2代细胞仍可保持腺细胞的生物学特征。Objective To establish the culture methods of submandibular gland cells from Wistar rats in vitro and lay a foundation for amplification of submandibular gland cells and gene therapy of salivary diseases. Methods The submandibular gland was obtained from Wistar rats which were one day old, after digested with LB3 and pancreatin, the submandibular gland cells were cultivated in DMEM/F12 culture fluid containing 2% fetal calf serum, epidermal gowth factor and insulin. The morphology of the culture and subculture cells was observed by phase contract microscope. The cell origin was identified by immunohistostaining. PAS staining was used to observe cell function of synthesis and secretion of polysaccharides. Results The morphous of primary cultured cells was round, triangle or polygon. At passage 2, the cells grew well, there was no change of morphous. The cells were positively stained with cytokeratin, E- cadherin and PAS, it indicated that the cells were epithelium origin and had the function of synthesis and secretion of potysaccharides. Conclusion The Wistar rat submandibular gland cells are successfully isolated and cultivated with enzyme digestion. The cultured submandibular gland cells at passage 2 still maintain their biocharacteristics.

关 键 词：颌下腺 细胞培养 细胞增殖 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R-332[医药卫生—基础医学]