犬乳腺上皮细胞的分离培养与鉴定

Culture and identification of mammary epithelial cells in canine milk

【目的】建立方便、经济的犬乳腺上皮细胞体外培养方法,用于犬乳腺癌的研究.【方法】以新产犬乳汁为材料,离心并收集乳汁中细胞,进行分离培养.用CCK-8法绘制细胞生长曲线,细胞角蛋白8免疫荧光染色鉴定上皮细胞.【结果和结论】试验材料培养后第3天即可见岛屿状的细胞克隆并伴有少量吞噬细胞.继续培养至单克隆细胞汇合后,可见细胞呈典型的石铺路状,单层平铺生长,且在细胞表层产生乳滴,细胞传代后吞噬细胞消失.CCK-8结果显示,细胞生长曲线呈"S"型.细胞免疫荧光结果显示,细胞角蛋白8呈阳性.表明从犬乳汁中可以分离培养出高纯度、生长迅速的犬乳腺上皮细胞.本试验建立了短时间内获得大量纯犬乳腺上皮细胞的方法.

【Objective】To establish an in vitro model of canine mammary epithelial cells for the study of the carcinogenesis and development of breast cancer. 【Method】 The cells were separated from canine milk and cultured. The cell growth curve was described using the CCK-8 assay. The expression of cytokeratin 8 was determined using cell immunofluorescence to identify mammary epithelial cell. 【Result and conclusion】 After cultured for 3 days, the cells were cloned with a shape like a typical island, and there were also a few phagocytes. Cultured for a few more days, the cells began to coalesce and form monolayers which could secrete lactogenesis drops. The phagocytes vanished after several passages. The cell growth curve was similar to a “S” shape. The immunofluorescent staining results showed that the cell of cytokeratin 8 was positive. The results indicated that the canine mammary epithelial cells separated from canine milk were highly pure and rapidly growing. This study established a model to isolate and culture massive canine mammary epithelial cells in a short time.