摘要
目的从MCF-7B乳腺癌细胞培养上清液中分离获得Exosomes,并进行形态学观察鉴定。方法通过低速、高速、超滤及超高速离心等方法,从MCF-7B乳腺癌细胞培养上清中分离Exosomes,用透射电子显微镜观察其形态特征,Lorry方法蛋白定量,通过用免疫印记进行组织相容性抗原(MHC)分子鉴定。结果MCF-7B乳腺癌细胞上清中分泌大量的Exosomes,电镜下观察呈椭圆形或圆形的双层膜的囊泡,直径为50～100nm,具有完整的细胞膜。结论用离心的方法可从MCF-7B乳腺癌细胞培养上清中提取Exosomes,有可能作为乳腺癌免疫治疗潜在的抗原,同时也为研究肿瘤的发病机制提供一个新的途径。
Objective To extract Exosomes from the cell culture supernatants of gastric cancer cell line MCF-7B. Methods MCF-7B cell lines were cultured and the ceil culture supernatants were collected to extract Exosomes with multiple centrifugation and ultrafiltration. Morphology of Exosomes was observed with transmission electron microscopy. Results Exosomes were purified from the MCF-7B cell culture supernatants. Exosomes exhibited heterogenous round or elliptic microvesicles ranging from 30 to 100 nm in diameter with intact membrane. Conclusion MCF-7B cell lines may secrete Exosomes, which could be used as potential specific antigens for immunotherapy against gastric cancer cells and might provide a new way to investigate the pathogenesis of cancer.
基金
辽宁省教育厅青年基金(05L544)
关键词
乳腺肿瘤
细胞系
细胞分离
Breast neoplasms
Cell line
Cell separation
