大鼠下颌下腺细胞培养及其生物学特性研究

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7 d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。

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

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 7 days 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) and α-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-8 and α-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.