利用膜過濾法分離卵巢癌細胞建立癌幹細胞系;Isolation of Ovarian Cancer Cells to Establish Cancer Stem Cells Using Membrane Filtration Method

NCUIR > Engineering College > National Central University Department of Chemical & Materials Engineering > Electronic Thesis & Dissertation > Item 987654321/94693

Please use this identifier to cite or link to this item: http://ir.lib.ncu.edu.tw/handle/987654321/94693

Title:	利用膜過濾法分離卵巢癌細胞建立癌幹細胞系;Isolation of Ovarian Cancer Cells to Establish Cancer Stem Cells Using Membrane Filtration Method
Authors:	蘇翊碩;SU, Yi-Shuo
Contributors:	化學工程與材料工程學系
Keywords:	卵巢癌;癌症幹細胞;尼龍膜;膜過濾法;聚乳酸-羥基乙酸共聚物/絲網;膜改質;Ovarian carcinoma;cancer stem cells;nylon membrane;membrane filtration method;poly(lactide-co-glycolic acid)/silk screen.;membrane modification
Date:	2024-07-25
Issue Date:	2024-10-09 15:24:22 (UTC+8)
Publisher:	國立中央大學
Abstract:	卵巢癌是女性生殖系統中最致命的惡性腫瘤之一，其發病率和死亡率一直引起全球範圍內的關注。其中，癌症幹細胞是一種在腫瘤中具有幹細胞特性的細胞，它們具有自我更新和分化成多種細胞型的能力。這些幹細胞被認為在癌症的發展和轉移過程中扮演著關鍵角色。因此，針對癌症幹細胞的治療策略可能有助於阻止腫瘤的再生和轉移，從而提高治療效果和預後。在這樣的背景下，本研究開發了一種膜過濾法。ES-2細胞株透過此方法，使用了不同的薄膜，分別是耐倫薄膜、聚乳酸-聚乙二醇酸薄膜和使用聚-L-賴氨酸與PVA-IA改質表面特性的的聚乳酸-聚乙二醇酸薄膜，透過改變薄膜的孔隙度度、厚度與表面性質達到純化CSCs的目的。純化效率將通過不同的檢測方式（1）流式細胞儀檢測CD44 & CD133的表達(2) 集落形成單位試驗。最後可以發現，在不同檢測方式的驗證下，透過與回收液和滲透液中的細胞相比，CSCs在遷移的細胞中具有更高的純度。接下來，為了測試膜過濾法將癌細胞從腫瘤細胞群分離的可行性，我們將卵巢癌細胞ES-2與人類成纖維細胞 CG1639共培養進行模擬。分別使用紅色與藍色的細胞追蹤劑染色來進行標記，以便利用流式細胞儀及螢光顯微鏡進行鑑定。結果顯示，在使用癌症幹細胞純化效率最高的10%聚乳酸-聚乙二醇酸薄膜時，膜遷移的ES-2細胞的純度有明顯的提升，表明薄膜上有更高比例的癌細胞殘留。這項技術顯示了作為分離癌症幹細胞（CSC）的可行策略的潛力，在即將到來的研究努力中具有精確醫學篩查的潛在應用。 ;Ovarian cancer is one of the most malignant tumors in the female reproductive system, and its incidence and mortality rate have been a global concern. Cancer stem cells (CSCs) are a type of cells in tumors that possess stem cell characteristics, with the ability to self-renew and generate tumors in organs or tissues in different sites. These stem cells are believed to play a crucial role in the development and metastasis of cancer. Therefore, treatment strategies targeting cancer stem cells may help preventing tumor regeneration and metastasis, thereby improving treatment outcomes and prognosis. A membrane filtration method was used to purify cancer stem cells using ovarian cancer cells in this study. ES-2 ovarian cancer cell line was used to be purified by membrane filtration method via several membranes including Nylon 11 membranes, poly(lactide-co-glycolic acid)/silk screen (PLGA/SK) membranes, and PLGA/SK membranes modified with poly-L-lysine or polyvinylalcohol-co-itaconic acid (PVA-IA) to achieve purification of CSCs by changing the porosity, thickness, and surface charges of the membranes. The purification efficiency was assessed using the following methods: (1) flow cytometry to detect the expression of CD44 & CD133 (CSC markers), and (2) colony-forming unit assay. The migrating cells after permeation through the membranes were found to have higher purity of CSCs compared to the cells in the recovery and permeate solutions. In order to evaluate the feasibility of separation of cancer cells from a tumor cell population using membrane filtration, ovarian cancer cells ES-2 were mixed with human fibroblast cells CG1639 to generate the simulated solution of primary tumor cell solution. The cell staining with Cell Tracker Red and Blue were used to identify ovarian cancer cells and fibroblasts, respectively using flow cytometry assay and fluorescence microscopy evaluation. The results showed that the purity of migrated ES-2 cells from 10% PLGA/SK membranes was the highest, indicating a high proportion of cancer cells remained on the membranes after the permeation of mixed solution of ovarian cancer cells and fibroblasts.
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