大鼠下颌下腺细胞培养及其生物学特性研究

A Study of Rat Submandibular Gland Cell Culture and Its Biological Characteristics

目的:体外培养大鼠下颌下腺细胞,并对其生物学特性进行研究,为涎腺疾病的探索提供体外模型。方法:无菌条件下取7d龄Wistar大鼠双侧下颌下腺,仔细分离脂肪、包膜、神经和血管,采用组织块培养法进行培养。运用酶消化法和差速贴壁法纯化细胞;免疫组化SP法检测Cytokeratin-8(CK-8)抗体和α-Amylase抗体的表达;PAS染色法检测细胞糖原分泌;扫描电镜(scanning electronic microscopy,SEM)观察细胞的形态学特征。结果:组织块培养法可成功获得下颌下腺细胞,免疫组化染色可见大部分细胞胞质为棕黄色,提示CK-8抗体表达阳性,α-Amylase抗体表达阳性;PAS染色可见胞质呈紫红色;SEM可见细胞伸出长短不一的伪足,胞核着色深,有些细胞可见分泌颗粒。结论:组织块培养法可以成功培养大鼠下颌下腺细胞,并且操作简便。

Objective：To culture submandibular gland cells of rats in vitro and study its biological characteristics so as to provide a cell model for salivary gland disease research.Methods：The bilateral submandibular glands were obtained from Wistar rats of 7days old in a sterile condition.Fat,capsule,nerve and blood vessel were carefully removed,followed by tissue-explant culturing.The cells were purified by enzymatic digestion and differential adhesion.The cell phenotype was immunohistochemically identified by cytokeratin-8（CK-8）andα-Amylase staining,and hepatin secreting ability of the cells was tested by PAS staining.Ultramicroscopic features of the cells was observed under the scanning electronic microscopy（SEM）.Results：Submandibular gland cells were successfully obtained by tissue-explant culturing.The cells gained were positively stained for both CK-8andα-Amylase.The cytoplasm manifested as purple red in PAS staining.Under SEM,cells had different lengths of pseudopodia and the nucleues were darker-stained;in some cells secretory granules were visible.Conclusion：Rat submandibular gland cells can be successfully cultured by the uncomplicated tissue-explant technique.