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  • 學位論文

密度梯度離心法純化外泌體之研究

Purification of Exosomes in Plasma via Density Gradient Centrifugation

指導教授 : 胡文聰
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摘要


外泌體為奈米外囊泡,粒徑大小約為30-150nm,,外層為脂質雙層膜構造,膜內攜帶多種訊號因子,包含蛋白質、醣類、核酸等,並傳遞訊號到其他細胞,扮演著細胞之間溝通的重要角色,由於大部分的細胞都可分泌外泌體,加上在許多體液中皆可發現外泌體的存在,因此成為非侵入式液態活檢的研究對象,近年來,許多文獻指出外泌體攜帶疾病相關的生物資訊,並且應用在疾病的診斷及追蹤。 本研究利用密度梯度離心法純化血液中的外泌體,結合疾病的生物標記,應用在退化性疾病上。研究結果提供血液中外泌體可以作診斷潛在生物標記之初步證據,實驗數據表示在疾病相關蛋白之表現,在統計上有一定的差異。所以,疾病蛋白之表現應該有潛力應用在檢測,然而需要更多的檢體量以驗證在臨床上的實際應用,期望未來能運用外泌體確實作為疾病之檢測。

並列摘要


Exosomes are nanosized vesicles with a diameter of 30-150 nm, encapsulated by lipid bilayer carrying proteins, lipids, and nucleic acids. The past decade has seen strong support that exosomes effectively deliver message to recipient cells and participate in cell-to-cell communication network. Furthermore, they have the ability to inherit various specific biological molecules from parent cells and reflect the pathological conditions when difference occurs in their composition. Recently, many studies indicated that exosomes have a relevant role in disease pathogenesis. In this study, we isolated exosomes by sucrose density gradient centrifugation and applied in neurodegenerative disease. The findings provided support for the notion that plasma-derived exosomes might perform as potential biomarkers of diagnosis in disease management. Future work requires further confirmation with additional clinical samples.

參考文獻


1.Hofmann, L., et al., The emerging role of exosomes in diagnosis, prognosis, and therapy in head and neck cancer. International journal of molecular sciences, 2020. 21(11): p. 4072.
2.Rufino-Ramos, D., et al., Extracellular vesicles: Novel promising delivery systems for therapy of brain diseases. Journal of Controlled Release, 2017. 262: p. 247-258.
3.Kalluri, R. and V.S. LeBleu, The biology, function, and biomedical applications of exosomes. Science, 2020. 367(6478): p. eaau6977.
4.Raposo, G. and W. Stoorvogel, Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol, 2013. 200(4): p. 373-83.
5.Lai, J.J., et al., Exosome Processing and Characterization Approaches for Research and Technology Development. Advanced Science, 2022: p. 2103222.

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