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姓名	邢永年(Yong-Nian Xing)  查詢紙本館藏	畢業系所	物理學系
論文名稱	結合基板引導膠原蛋白自組裝法與利用紫外光曝光及胰蛋白酶蝕刻產生微米圖案的技術來製作膠原蛋白纖維島嶼以應用於細胞表現型之控制 (Control of cell phenotype by collagen fibril islands fabricated by substrate-guided self-assembly of collagen molecules and micro-patterning using masked UV exposure and trypsin etching)
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摘要(中)	研究顯示，微米及奈米尺度的基板會影響貼附在上面的細胞，包括貼附形狀、遷移、繁殖、分化等等。細胞外基質像膠原蛋白纖維這種具有特殊結構的基板在控制細胞的表現型上面就扮演了重要的角色。在本實驗中，我們結合自組裝膠原蛋白分子和微米圖案光刻製造膠原蛋白纖維島嶼，我們把人類間充質幹細胞(MSC)養在膠原蛋白纖維島嶼基板上面，觀察細胞是否會貼附在島嶼上面並改變自身形狀，並在基板製作時滴入PLL-g-PEG這種聚合物，之後觀察細胞貼附在島嶼上的情形分析PLL-g-PEG對基板的影響，藉由觀察細胞貼附的狀況來改善PLL-g-PEG的參數，最終達到完全控制細胞的貼附形狀。未來，我們會加入不同的分化劑來觀察MSC貼附在不同的膠原蛋白纖維島嶼上的分化速度，再用共厄焦顯微鏡來觀察細胞內的變化，像是用DNA、F-atin等等，利用觀察的結果改變膠原蛋白纖維島嶼的形狀，讓MSC分化的速度到達最快。
摘要(英)	It has been known that the microscale and nanoscale substrate topography of cell-culturing substrate can regulate cell adhesion, migration, proliferation, and differentiation. It is also known that extracellular matrix is made of most importantly collagen fibrils, which possess specific molecular identity and structural characteristics that may be crucial for controlling cell behaviors such as cell phenotype. In this work, we combined the techniques of self-assembly of collagen molecules and lithographic micropatterning to produce collagen fibril islands. The collagen fibril island substrates were used to culture human mesenchymal stem cells (MSC) to control their shape. We also added PLL-g-PEG on the substrate and observed the cell type to know PLL-g-PEG contribution. In future, We will observe MSC differentiation rate by various collagen fibril islands.We will use DNA, F-actin to observe cell internal transform by collagen fibril islands.
關鍵字(中)	★ 膠原蛋白 ★ 微米圖案 ★ 幹細胞 ★ 原子力顯微鏡	關鍵字(英)	★ Collagen ★ Micro-patterning ★ MSC ★ AFM
論文目次	Abstract IV 摘要 V 致謝 VI 目錄 VII 圖目錄 IX 一.緒論 1 1.1表觀遺傳學 1 1.2細胞外基質 1 1.3膠原蛋白纖維 1 1.4控制細胞貼附大小影響細胞分化 5 1.5 PLL-g-PEG鈍化基板表面 10 1.6 膠原蛋白的光分解 11 二.實驗材料與方法 13 2.1 膠原蛋白薄膜製備 13 2.2 膠原蛋白纖維島嶼製程 14 2.3 強化細胞對膠原蛋白島嶼的貼附性 15 2.4 間充質幹細胞的培養 16 2.5 光蝕刻裝置檢測 16 2.6 膠原蛋白微米圖案檢測 18 2.7 膠原蛋白螢光染色 19 2.8 DAPI與Phalloidin螢光染色 19 2.9 引導MSC分化成脂肪細胞 19 2.10 Oil red染色流程 20 三.結果 21 3.1曝光架設的改進 21 3.2 膠原蛋白薄膜與島嶼 21 3.3膠原蛋白的熱處理(本實驗由Saitong Muneekaew 提供數據) 23 3.4 PLL-g-PEG對膠原蛋白纖維島嶼的影響 24 3.5 MSC在不同PLL-g-PEG時間下的影響 25 3.6引導MSC分化成脂肪細胞 29 四.結論與未來展望 31 4.1 結論 31 4.2未來展望 31 五.參考文獻 33
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指導教授	陳賜原(Szu-yuan chen)	審核日期	2019-1-29
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