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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 侯詠德 | |
| dc.contributor.author | Yi-yu Xu | en |
| dc.contributor.author | 許乙雨 | zh_TW |
| dc.date.accessioned | 2021-06-15T12:35:47Z | - |
| dc.date.available | 2017-08-24 | |
| dc.date.copyright | 2016-08-24 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-07-30 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50307 | - |
| dc.description.abstract | 為了克服器官捐贈的不足以及肝切除手術對病患所造成的負擔，本研究擬開發使受損肝臟的肝細胞恢復的肝臟貼片。肝臟貼片是利用肝組織工程(Liver tissue engineering)的脫細胞化技術所得到的肝臟脫細胞外間質(Decellularized liver ma-trix; DLM)作為本體，並將肝細胞生長因子(Hepartocyte growth factor, HGF)/肝素(Heparin)複合體修飾固定於DLM上。實驗結果得知: (1) Heparin固定於DLM film上的密度約為30 μg/cm2；(2)培養三天後，正常肝細胞在HGF/heparin-immobilized DLM的培養條件下，細胞相對生存率(Relative cell viability)提升20—30 %，肝蛋白(Albumin)分泌量可提升約20 %；(3)經過半乳糖胺(D-galactosamine)毒化處理後的肝細胞在HGF/Heparin-immobilized DLM的培養條件下，Lactate Dehydrogenase (LDH) 的活性為10 milliunits/mL，比起negative control組(毒化後的肝細胞在normal dish上繼續培養)的培養條件，損傷程度只有原本的一半，並且肝細胞機培養三天後，肝細胞能夠恢復至與無毒化組別的相同水準。本研究證明了HGF/Heparin-immobilized DLM film不但具有提升肝細胞活性、且提升肝蛋白的分泌量，另外更能夠恢復受到毒化後的肝細胞的活性並延緩肝細胞毒害的效果。這個結果對於未來製造出具備修復肝臟器官的生物貼片材料，具有十分重要的意義。 | zh_TW |
| dc.description.abstract | In order to overcome the shortage of organ donation and injury of patients after partial hepatectomy (PH) treatment, we aimed at developing a liver film for hepato-cytes regeneration through liver tissue engineering. In this study, a HGF/heparin-immobilized decellularized liver matrix film (HGF/heparin complex coated on DLM film) was developed for hepatocytes regeneration in liver injury. In the result, (1) the amounts of immobilized heparin on DLM film was 30 μg/cm2 when the initial heparin concentrations were 1 mg/mL. (2) The relative cell viability and albu-min synthesis of the hepatocytes on HGF/heparin-immobilized DLM film was 20-30 % and 20 % superior than on normal dish at 3 days of culture, respectively. (3) The lactate dehydrogenase activity of the D-galactosamine-induced injury of hepatocytes on hepa-rin-immobilized DLM film cultures was 10 milliunits/mL, which was 50 % lower than that in the D-galactosamine-induced injury of hepatocytes on normal dish cultures, and albumin synthesis can recover to the same level as non-toxic hepatocytes cultured on normal dish. The HGF/heparin-immobilized DLM film showed highly potential in maintaining hepatocyte culture and also in repairing injured hepatocytes from D-galactosamine. It is believed that this HGF/heparin-immobilized DLM film has promising potential for hepatocyte transplantation, and could be applied for future use in liver tissue engineering. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T12:35:47Z (GMT). No. of bitstreams: 1 ntu-105-R03631045-1.pdf: 3894708 bytes, checksum: 314aca6e7a1bb52ff86e191fb0e6dc15 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 致謝 i
摘要 ii Abstract iii 目錄 iv 圖目錄 vii 表目錄 ix 第一章 前言 1 1.1 背景 1 1.2 研究目的 2 1.3 實驗架構 3 第二章 文獻探討 4 2.1 再生醫學 4 2.1.1幹細胞技術 4 2.1.2組織工程 5 2.2 肝組織工程 6 2.2.1肝組織工程相關生長因子 7 2.2.1.1 Hepatocyte Growth Factor 7 2.2.1.2 Heparin 10 2.2.1.3 Heparin-HGF複合體 12 2.2.2肝組織工程相關生物材料 13 2.2.2.1 Collagen 13 2.2.2.2細胞外間質與肝臟脫細胞化間質 16 2.3 脫細胞化技術 19 2.3.1脫細胞化試劑/方法 21 2.3.2肝臟脫細胞化方法 22 2.4受損肝臟的自體恢復機制 23 2.5 D-galactosamine毒化機制 25 第三章 材料與方法 27 3.1 實驗藥品、耗材、儀器設備與實驗動物 27 3.1.1實驗藥品 27 3.1.2實驗耗材 27 3.1.3實驗儀器設備 28 3.1.4實驗動物 28 3.2脫細胞化肝臟間質製備 29 3.3 DLM Film的製備 30 3.4 Heparin-immobilized DLM film製備 31 3.5 HGF/heparin-immobilized DLM film製備 32 3.6 DLM film用於肝細胞培養 33 3.7 DLM film用於半乳糖胺D-galactosamine誘導毒化後肝細胞的培養 34 3.8 實驗相關檢測方法 35 3.8.1 Heparin濃度檢測 35 3.8.2 HGF濃度檢測 35 3.8.3粒腺體活性檢測 36 3.8.4 Albumin分泌量檢測 36 3.8.5 LDH活性檢測 37 3.8.6 肝細胞DNA含量檢測 37 第四章 結果與討論 38 4.1脫細胞化肝臟間質製備最佳化條件的探討 38 4.2脫細胞化肝臟間質的製備結果 40 4.3 DLM film製備最佳化條件之探討 41 4.4 Heparin於DLM Film上的固定化量探討 43 4.5 HGF於heparin-immobilized DLM Film上的固定化量探討 45 4.6 DLM film用於肝細胞培養的探討 46 4.7 D-galactosamine毒化肝細胞的條件探討 48 4.8 DLM film用於半乳糖胺D-galactosamine誘導毒化後肝細胞培養的探討 49 第五章 結論與未來展望 58 5.1結論 58 5.2展望 59 參考文獻 60 | |
| dc.language.iso | zh-TW | |
| dc.title | 肝細胞生長因子與肝素固定化的脫細胞化肝臟間質應用於肝組織工程的研究 | zh_TW |
| dc.title | HGF/Heparin-immobilized Decellularized Liver Matrix for Liver Tissue Engineering | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳力騏,黃祥博 | |
| dc.subject.keyword | 脫細胞化肝臟間質,肝細胞生長因子/肝素複合體,肝臟貼片,肝細胞再生,肝組織工程, | zh_TW |
| dc.subject.keyword | decellularized liver matrix,HGF/Heparin complex,liver film,hepatocytes regeneration,liver tissue engineering, | en |
| dc.relation.page | 71 | |
| dc.identifier.doi | 10.6342/NTU201601159 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-08-01 | |
| dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
| dc.contributor.author-dept | 生物產業機電工程學研究所 | zh_TW |
| 顯示於系所單位： | 生物機電工程學系 | |
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