牦牛支气管上皮细胞的分离培养与鉴定

Isolation,culture and identification of yakbronchial epithelial cells

为了分离培养牦牛支气管黏膜上皮细胞并传代培养,利用支气管结扎灌注酶冷消化法和支气管组织贴壁法分离培养牦牛支气管黏膜上皮细胞并传代培养,第二代细胞制作细胞爬片通过H.E.染色,扫描电镜和免疫荧光分别对培养的上皮细胞的形态结构进行鉴定,结果两种方法都获得了牦牛支气管黏膜上皮细胞,细胞活性好,纯度高;H.E.染色可见细胞形态呈梭形、圆形、多角形等,细胞核深染,细胞质淡红色;扫描电子显微镜检测中,明显可见细胞的纤毛;免疫荧光鉴定细胞核清晰可见,细胞核呈亮绿色,表明分离培养的细胞为上皮细胞,可进一步为牦牛支气管黏膜上皮细胞粘附模型的建立提供材料和奠定技术基础。

In order to isolate and culture the bronchial epithelial cells of yak, the bronchial mucosal epithelial cells wereisolated and cultured by the method of bronchial ligation and cold digestion and bronchial tissue adherence. The second generation cells were observed by H.E. staining, and the morphology and structure of cultured epithelial ceils were identified by scanning electron microscopy and immunofluorescence, results show that the two methods were used to obtain the yak bronchial epithelial cells, which with high activity and high purity ; H.E. staining showed that the cultured ceils were spindle shaped, round, polygonal, hyperchromatic nuclei, and cytoplasm pale red. Scanning electron microscopy showed that the cells were clearly visible. Fluorescent identification showed that the nucleus was clearly visible and the nucleus was bright green. The results showed that the cultured cells were epithelial cells.