Purification and functional study of lung cancer cell exosomes

• Main Authors: LIU,KANG-YU, 劉康宇
• Other Authors: HUANG, SHU-LING
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/g5s294

碩士 === 國立聯合大學 === 化學工程學系碩士班 === 107 === Exosomes are membrane-like-extracellular vesicles with diameter between 20-200nm secreted by most of the eukaryotic cells, which are suggested to have drug resistance effects on cancer cells. Despite a considerable amount of exosomes that could be found in biological medium, the extraction and purification process remains challenging, the high cost and low yield of commercially available kits have limited the applications and development. Here, a novel, affordable and scalable platform was successfully established to effectively purify exosomes from biological medium via precipitation, while preserving the molecular functions of that. In this study, the quality of extraction and purification of exosomes secreted by non-small cell lung cancer PC9 cells were investigated between ExoQuick-TC™, a commercial kits, and polyethylene glycol (PEG) with various molecular weights (Mn), concentrations and pH. Particle size of exosomes purified by all methods were similar, while four-folded yield as well as increased expression of marker protein of exosomes were obtained via 14% PEG with Mn of 8000 at pH 5.4 when compared with ExoQuick-TC™. Besides, PC9 cells pre-treated with exosomes purified by this platform had resulted in 3-5 times increased expression of anti-drug related proteins, 25% reduced apoptosis rate as well as 15% increased cell viability when treated with an anti-cancer drug, gefitinib, indicating that the molecular functions of exosomes purified in this study were preserved.