目的分离培养人乳腺肿瘤原代细胞,为研究人乳腺肿瘤的病理研究提供实验模型。方法采用Ⅲ型胶原酶消化法和差速离心法分离培养人乳腺肿瘤细胞,对存活乳腺肿瘤细胞进行细胞形态观察和免疫荧光染色鉴定。结果乳腺肿瘤细胞接种24 h后开始贴壁,培养8~10 d生长融合成片,在倒置显微镜下观察,细胞呈上皮细胞样,细胞角蛋白(CK)-19免疫荧光染色呈阳性,细胞纯度达到95%以上。结论采用本法体外分离培养人乳腺肿瘤原代细胞,细胞生长良好,纯度高,可用于人乳腺肿瘤病理等方面研究。
Objective To isolate and culture human breast cancer primary cells, and to provide experimental model for pathological study of human breast cancer. Methods Human breast cancer cells were digested by collagenase Ⅲ and isolated by differential centrifugation. The survival breast cancer cells were determined by cytomorpholo-gy and identified by immunofluorescent staining. Results The breast cancer cells became adherent at 24 h after inoculation, and grew to confluence at 8-10 d after culture. Under inverted microscope, epithelioid-like cells were found.Immunofluorescent staining for cytokeratin 19 showed positive reaction, and the purity of the cells was more than95%. Conclusion The isolated and cultured human breast cancer primary cells in this study grow well with high purity, which can be used in the pathological studies of human breast cancer.