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研究生： 余芮萍
Jui-Ping Yu
論文名稱： 研究複合水凝膠在細胞生物上的應用
The design of composite hydrogels for cell biology applications
指導教授： 黃人則
Jen-Tse Huang
何明樺
Ming-Hua Ho
口試委員: 黃人則
Jen-Tse Huang
何明樺
Ming-Hua Ho
游佳欣
Jiashing Yu
學位類別： 碩士
Master
系所名稱： 工程學院 - 化學工程系
Department of Chemical Engineering
論文出版年： 2022
畢業學年度： 110
語文別： 中文
論文頁數： 67
中文關鍵詞： 生物蛋白 、胜肽 、自組裝分子 、水凝膠 、細胞支架
外文關鍵詞： organism proteins, peptides, self-assembly, hydrogels, cell scaffolds
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    • 摘要 i ABSTRACT ii 致謝 iii 目錄 iv 圖目錄 vii 表目錄 viii 附圖目錄 ix 第ㄧ章 緒論 1 1.1 貽貝中的自組裝蛋白 1 1.2 自組裝分子的性質與應用 3 1.2.1 自組裝分子間之相互作用力及調控 3 1.2.2 自組裝分子之結構 3 1.3 胜肽水凝膠的應用 4 1.3.1 胜肽水凝膠之可注射性質 4 1.4 三維細胞支架 5 1.5 研究目的 7 第2章 實驗儀器與材料 8 2.1 實驗儀器列表 8 2.2 實驗材料 10 2.2.1 胜肽合成與純化試劑配置 10 2.2.2 電子顯微鏡樣品製備材料 10 2.2.3 細胞材料 11 2.2.2 藥品表 11 第3章 實驗方法與使用儀器介紹 13 3.1 胜肽樣品製備 (合成、純化與鑑定) 13 3.1.1 固相胜肽合成法 (Solid-Phase Peptide Synthesis, SPPS) 13 3.1.2 高效液相層析 (High-Performance Liquid Chromatography, HPLC) 14 3.1.3 基質補助雷射脫附游離質譜法 (MALDI Mass Spectrometry) 15 3.2 聚多巴胺樣品製備 15 3.2.1 多巴胺的聚合 15 3.2.2 螢光光譜法之鑑定 (Fluorescence Spectrometry) 16 3.3水凝膠材料性質分析 17 3.3.1 流變儀 (Rheometer) 17 3.3.2 穿透式電子顯微鏡 (Transmission Electron Microscopy, TEM) 18 3.3.2.1 胜肽纖維負染步驟 18 3.3.3 掃描電子顯微鏡 (Scanning Electron Microscope, SEM) 19 3.3.3.1 水凝膠樣品鍍膜步驟 19 3.3.4 水凝膠之自我修復能力 (Self-healing) 19 3.4 三維細胞支架 (Three-dimensional scaffold) 之分析 20 3.4.1 三維細胞支架 (Scaffold) 的製備 20 3.4.2 細胞培養和繼代 20 3.4.3 螢光染色 (Fluorescence staining) 21 3.4.4 共軛焦顯微鏡 (Confocal Microscope) 22 第4章 實驗結果與討論 23 4.1胜肽自組裝水凝膠材料的設計 23 4.1.1 貽貝蛋白之序列挑選 23 4.1.2 置換不同胺基酸組成的胜肽 24 4.2 胜肽材料的合成 24 4.2.1胜肽之性質純化和鑑定 25 4.3 水凝膠的製備 25 4.3.1 水凝膠在不同pH值下的差異 26 4.3.2 鹽離子有助於形成水凝膠 28 4.3.3 胜肽序列之等電點 29 4.3.4 以流變儀鑑定材料的流動性質 30 4.3.4.1 不同濃度下的測量 30 4.3.4.2 不同 pH 下的測量 31 4.3.4.3 不同胜肽序列下的測量 32 4.3.4.4 水凝膠材料自修復性質 33 4.4 以電子顯微鏡觀察水凝膠 34 4.4.1 穿透式電子顯微鏡下之水凝膠 34 4.4.2 掃描式電子顯微鏡下之水凝膠 35 4.5 混合聚多巴胺以增強水凝膠性質 36 4.5.1多巴胺的聚合 36 4.5.1.1 聚多巴胺的合成 36 4.5.1.2 聚多巴胺之螢光光譜 37 4.5.2 胜肽水凝膠混合不同濃度的聚多巴胺 37 4.5.3 水凝膠材料可注射性質 38 4.6 水凝膠的膨潤比 39 4.7 測試纖維母細胞在水凝膠上的生長 39 4.7.1不同成膠條件下對培養基pH的影響 39 4.7.2 纖維母細胞在胜肽水凝膠上之貼附 40 4.7.2.1 以正立式顯微鏡觀察水凝膠表面 40 4.7.2.2 水凝膠作為三維細胞支架 (Scaffold) 之研究 41 4.7.2.3 水凝膠的孔洞率對細胞貼附之影響 42 4.7.3 降低聚多巴胺濃度對水凝膠之影響 44 4.7.3.1 不同聚多巴胺濃度下之流變性質 44 4.7.3.2 不同聚多巴胺濃度下之水凝膠孔洞率 45 4.7.3.3 細胞在水凝膠孔洞上的分佈 45 第5章 總結 47 附圖 48 參考資料 54

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