摘要
目的探讨人类肥大乳房乳腺上皮细胞的原代培养方法。方法采用胶原酶消化培养法，在体外进行人类肥大乳房乳腺上皮细胞的分离培养，用倒置显微镜观察、细胞爬片、HE染色和细胞角蛋白免疫组织化学染色的方法对分离培养的细胞进行形态学观察和鉴定。结果倒置显微镜下观察细胞形态呈鹅卵石型或多角型，部分形态不规则，增殖的过程中形成岛屿状闭合型的细胞群，细胞之间连接紧密。细胞HE染色可见细胞胞质呈粉红色或浅紫色，细胞核呈蓝紫色，圆形或椭圆形，核中可见深蓝色的染色体。免疫组化鉴定结果显示，培养的细胞胞浆内可见棕黄色的细胞角蛋白着色，表达上皮细胞特异的细胞角蛋白18。结论应用酶消化法和条件培养液可以获得纯度较高的人类肥大乳房乳腺上皮细胞。
Objective To investigate the primary culture of mammary epithelial cells derived from human breast hypertrophy. Methods Mammary epithelial cells of human breast hypertrophy were isolated and cultured by the collagenase digestion method. The morphology observation and identification of the cultured cells were performed by inverted microscope observation, HE staining and cytokeratin immunohistochemical staining. Results Most of the cultured cells were pebbles-like or polygon under the inverted microscopy. Some had irregular form. They had typical island-like appearance during multiplication with close connection between the cells. The HE staining results showed the cytoplasm was stained pink or lilac, the nucleus was stained bluish violet and was round or oval in shape, with clearly visible chromosomes in dark blue. The cytokeratin immunohistochemical staining demonstrated the tissue-specific expression of cytokeratin 18 in epithelial cells by the cytoplasm stained claybank. Conclusion High purity of primary mammary epithelial cells derived from human breast hypertrophy can be obtained by the collagenase digestion method and conditioned medium.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2012年第5期356-359,共4页
Chinese Journal of Plastic Surgery
关键词
乳房肥大
上皮细胞
细胞培养技术
Hypermastia
Epithelial cell
Cells culture techniques
