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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 楊台鴻 | |
dc.contributor.author | Chun-Hung Lin | en |
dc.contributor.author | 林俊宏 | zh_TW |
dc.date.accessioned | 2021-06-15T02:54:27Z | - |
dc.date.available | 2009-08-14 | |
dc.date.copyright | 2009-08-14 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-08-03 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/44384 | - |
dc.description.abstract | 本實驗是以乾式法將高分子乙烯乙烯醇(EVAL)聚合物製成薄膜之後,用化學改質的方式將表面氫氧基(-OH)接枝甲基丙烯醯氯(MAC)以間位氯化過氧苯酸(mCPBA)將烯烴雙鍵進行環氧化反應化後接枝離胺酸(L-lysine)單體(E-Lys)和1,4-丁二胺(E-14)。並且將接枝離胺酸(L-lysine)後的薄膜用碳化二亞胺(EDC)修飾離胺酸上的羧酸基接上14-丁二胺(E-Lys-14)。
之後利用體外細胞培養的方法,來探討材料與神經細胞之間的生物適合性,將改質前以及改質後的薄膜培養小鼠初代顆粒型神經細胞,以MTT測試來比較細胞的活性並且用光學顯微鏡和曠時攝影來觀察細胞貼附在接枝1,4-丁二胺、離胺酸以及經1,4-丁二胺修飾之離胺酸並和塗佈聚離胺酸(poly-D-lysine)的乙烯乙烯醇薄膜之細胞型態與行為。 另外再利用螢光顯微鏡來觀察神經細胞是否有發展出功能性之結構。最後利用西方點墨法與麩胺酸釋放來測量神經細胞在各材料上之蛋白質表現量與神經傳導功能之差異。 整體而言小鼠顆粒型神經細胞培養於離胺酸修飾1,4-丁二胺時(E-Lys-14),不論是細胞活性、整體蛋白質表現,甚至是培養天期都優於EVAL塗佈聚離胺酸薄膜。接枝離胺酸修飾1,4-丁二胺的確具有取代塗佈聚離胺酸培養神經細胞的潛力。 | zh_TW |
dc.description.abstract | We prepare poly(ethylene-co-vinyl alcohol)(EVAL) membranes by the dry processing ,and then grafted methacryloyl chloride (MAC)on the hydroxyl group of these membrane surface by the way of chemical modification. Furthermore, we grafted different diamines, lysine and modified lysine with 1,4-diaminobutane by EDC/NHS method on them.
We cultured the cerebellar granule neurons of the Wistar rat on the modified membranes and compared with the relative activity by MTT assays. In addition, the cell morphology and behavior was observed by optical microscope and time-lapse. The functional structures of cerebellar granule neurons cultured on different membranes were observed by fluorescence microscope. We also used Western blot and glutamate release to mesure the protein expression and neural transmission. Overall, the modification of the EVAL with lysine modified 1, 4-diaminobutane ( E-Lys-14) have better relative activity, higher protein expression level and longer culture duration than coating PDL. E-Lys-14 may be the batter substrate to substitude coating PDL to culture neurons. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T02:54:27Z (GMT). No. of bitstreams: 1 ntu-98-R96549017-1.pdf: 5480297 bytes, checksum: 094d2a2a00e84aba8725b293a533d96d (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 第1章 緒論 1
第2章 文獻回顧 4 2-1高分子薄膜 4 2-1-1高分子薄膜定義 4 2-1-2高分子薄膜結構 4 2-1-3薄膜的製備 4 2-2神經細胞培養 5 2-2-1神經系統簡介 6 2-2-2神經細胞突觸之形成 7 2-2-2培養神經細胞用途 8 2-2-3影響神經細胞生長因素 9 2-2-4神經細胞之電位傳導與神經傳導物質之釋放 11 2-3薄膜改質 13 2-3-1改質簡介 13 2-3-2材料表面改質與細胞生長的關係 14 第3章 實驗材料與方法 16 3-1實驗架構 16 3-2試藥原料 17 3-3儀器 20 3-4試藥配置 25 3-5薄膜製備 30 3-5-1 EVAL薄膜製備 30 3-5-2薄膜表面改質 30 3-6神經細胞培養 35 3-6-1薄膜前處理 35 3-6-2塗佈聚離胺酸 35 3-6-3初代細胞培養(primary cell culture)步驟 35 3-6-4 MTT測試評估神經細胞生長情形 36 3-6-5 Time-lapse觀察 37 3-7 塗佈聚離胺酸4小時、7、14、21天之釋放量 37 3-8 LDH測試評估細胞對材料之貼附能力 38 3-9免疫螢光染色 (immunofluorescence stain) 38 3-10西方點墨法 (Western blot) 39 3-11神經細胞功能測試 41 3-11-1 突觸素I蛋白 (Phospho-Synapsin I) 磷酸化程度評估 41 3-11-2 穀胺酸(Glutamate)釋放 41 第4章 結果與討論 43 4-1 EVAL接枝1,4-丁二胺、離胺酸、離胺酸修飾1,4-丁二胺之細胞活性MTT測試 43 4-2 神經細胞在不同材料的光學顯微鏡照片 45 4-3 神經細胞在不同材料的突觸長度 47 4-6 聚離胺酸釋放之測試 47 4-4神經細胞在不同材料的無血清培養光學顯微鏡照片 48 4-5 神經細胞在不同材料的曠時攝影觀察 49 4-7 神經細胞在不同材料上有血清與無血清之貼附強度比較 50 4-8 EVAL塗佈10% FBS培養基對神經細胞行為之影響 50 4-9 神經細胞在不同材料的螢光染色照片 51 4-10 神經細胞在不同材料的蛋白質表現與定量 52 4-11神經細胞在不同材料的功能性評估 54 第5章 結論 59 第6章 參考文獻 60 | |
dc.language.iso | zh-TW | |
dc.title | 聚乙烯乙烯醇薄膜接枝離胺酸與1,4-丁二胺對神經元細胞行為與功能性之影響 | zh_TW |
dc.title | Behavior and Functionality of Neurons Cultured on EVAL Membranes Modified With Lysine and 1,4-Diaminobutane | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 孫一明,楊銘乾 | |
dc.subject.keyword | 聚乙烯乙烯醇,小腦顆粒神經細胞,甲基丙烯醯氯,碳化二亞胺,1,4-丁二胺,離胺酸,聚離胺酸, | zh_TW |
dc.subject.keyword | EVAL,cerebellar granule cells,methacryloyl Chloride,EDC,PDL,1,4-diaminobutane,lysine, | en |
dc.relation.page | 107 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-08-04 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 高分子科學與工程學研究所 | zh_TW |
顯示於系所單位: | 高分子科學與工程學研究所 |
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