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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50678完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊台鴻(Tai-Horng Young) | |
| dc.contributor.author | Tzung-Yu Chen | en |
| dc.contributor.author | 陳宗郁 | zh_TW |
| dc.date.accessioned | 2021-06-15T12:52:13Z | - |
| dc.date.available | 2026-07-19 | |
| dc.date.copyright | 2016-08-02 | |
| dc.date.issued | 2016 | |
| dc.date.submitted | 2016-07-19 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/50678 | - |
| dc.description.abstract | 本研究分別從人類膝關節中的滑膜組織及前十字韌帶分離出細胞，進一步將其放置在幾丁聚醣上進行培養，培養幾天後觀察細胞的增生及老化表現的改變。
幾丁聚醣是一個天然、生物可分解、生物可相容性、無生物毒性且經由美國食品藥品管理局認可的多醣類，本研究使用其作為培養細胞之基材。當前十字韌帶與滑膜細胞分別種植於幾丁聚醣基材上時，細胞都會呈現懸浮且聚集的型態。讓細胞在這個懸浮的狀態下持續三天後，再回種到TCPS (Tissue culture polystyrene) 上，並以沒有做任何處理的細胞作為對照組，探討細胞型態及功能的改變。 接著，我們從兩個層面來探討細胞功能，第一是增生能力，我們建立細胞生長曲線、運用MTT ([3-[4,5-dimethylthiahiazo-2-yl]-2,4-dipheny- tetrazolium bromide]) assay和BrdU (5-bromo-2’-deoxiuridine) 生長偵測組做ELISA定量，互相佐證其細胞增生能力的改變。第二則是老化表現，我們運用Senescence-associated --galactosidase (SA--gal) 染色的方法來觀察SA--gal活性是否有被抑制。 最後，我們蒐集各種不同年齡病人檢體所獲得的細胞，進而探討本細胞處理方法對於不同年齡病人間的效果。 | zh_TW |
| dc.description.abstract | In this study, we isolated primary cells form human synovial membrane and human anterior cruciate ligament. First, we seed the cells on the chitosan substrate and culture for a few days. After this treatment, we studied the change of cell function.
Chitosan is a natural, biodegradable, biocompatible, non-toxic and U.S. Food and Drug Administration (FDA) approved polysaccharide. In this study, chitosan was used as the coating substrates. When synovial cells and anterior cruciate ligament fibroblasts were cultured on chitosan, all cells became suspended and aggregated into spheroids. After three days, we seeded the cell spheroids on TCPS (Tissue culture polystyrene), and studied change of cell proliferation and cell senescence expression. Furthermore, we established the population doubling curve, used MTT ([3-[4,5-dimethylthiahiazo-2-yl]-2,4-dipheny- tetrazolium bromide]) assay and BrdU (5-bromo-2’-deoxiuridine) assay, to study the cell proliferation change. We used the inverted microscope to observe the cell morphology and used senescence-associated-β-galactosidase (SA-β-gal) to study the cell senescence expression. Finally, we collected cells from various patients to study the relationship between the effect of chitosan treatment and patient’s age. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T12:52:13Z (GMT). No. of bitstreams: 1 ntu-105-R03548011-1.pdf: 2461835 bytes, checksum: bc6a721ef42bd25f9785b705057f4f02 (MD5) Previous issue date: 2016 | en |
| dc.description.tableofcontents | 目錄
摘要 i Abstract ii 目錄 iii 圖目錄 vi 第一章 緒論 1 第二章 文獻回顧 2 2.1 膝關節（keen joint） 2 2.2 前十字韌帶（anterior cruciate ligament, ACL） 2 2.3 滑膜（synovial membrane） 2 2.4 老化（aging） 3 2.5 幾丁聚醣（chitosan） 5 第三章 實驗材料與方法 6 3.1 實驗架構 6 3.2 實驗材料 7 3.3 儀器 9 3.4 試劑配製 12 3.5 實驗方法 14 3.5.1 幾丁聚醣薄膜製備(Preparation of chitosan substrates) 14 3.5.2 細胞培養(Cell culture) 14 3.5.3 幾丁聚醣處理(Chitosan Treatment) 15 3.5.4 雙重幾丁聚醣處理(Double Treatment) 15 3.5.5 細胞繼代曲線(Population doubling curve) 16 3.5.6 MTT assay 17 3.5.7 BrdU assay 17 3.5.8 SA-β-gal staining 18 3.5.9 不同病人之細胞BrdU生長偵測與SA-β-gal活性定量關係圖 18 3.5.10 統計分析 19 第四章 實驗結果 20 4.1 初代細胞培養（primary cell culture） 20 4.1.1 滑膜細胞(SCs) 20 4.1.2前十字韌帶纖維母細胞(ACLs) 20 4.2 細胞於TCPS及幾丁聚醣（chitosan）基材上的型態 20 4.3 細胞繼代曲線（population doubling curve, PD curve） 20 4.4 細胞繼代次數的最大值(Maximum of population doubling times) 21 4.5 細胞培養天數與SA-β-gal活性 22 4.6 幾丁聚醣處理後的細胞型態（Morphology） 22 4.7 幾丁聚醣處理後的繼代曲線 22 4.8 細胞增生試驗（BrdU生長偵測） 23 4.9 細胞活性試驗（MTT assay） 24 4.10 細胞老化表現（SA-β-gal staining） 25 4.11 BrdU生長偵測與病人年齡 26 4.12 SA-β-gal staining與病人年齡 26 4.13 雙重幾丁聚醣處理 27 第五章 討論 28 5.1 初代細胞培養 28 5.2 細胞於TCPS及幾丁聚醣基材上的型態 28 5.3 細胞繼代曲線 28 5.4 細胞繼代次數的最大值 29 5.5 細胞培養天數與SA-β-gal活性 29 5.6 幾丁聚醣處理後的細胞型態 30 5.7 幾丁聚醣處理後的繼代曲線 30 5.8 細胞增生試驗 31 5.9 細胞活性試驗 31 5.10 細胞老化表現 32 5.11 BrdU生長偵測與病人年齡 32 5.12 SA-β-gal staining與病人年齡 33 5.13 雙重幾丁聚醣處理 33 第六章 結論 35 第七章 參考文獻 36 第八章 圖表 40 | |
| dc.language.iso | zh-TW | |
| dc.subject | 幾丁聚醣 | zh_TW |
| dc.subject | 前十字韌帶纖維母細胞 | zh_TW |
| dc.subject | 幾丁聚醣 | zh_TW |
| dc.subject | 細胞增生 | zh_TW |
| dc.subject | 老化表現 | zh_TW |
| dc.subject | 細胞增生 | zh_TW |
| dc.subject | 滑膜細胞 | zh_TW |
| dc.subject | 前十字韌帶纖維母細胞 | zh_TW |
| dc.subject | 老化表現 | zh_TW |
| dc.subject | 滑膜細胞 | zh_TW |
| dc.subject | chitosan | en |
| dc.subject | synovial cells | en |
| dc.subject | anterior cruciate ligament fibroblasts | en |
| dc.subject | cell proliferation | en |
| dc.subject | senescence expression | en |
| dc.subject | chitosan | en |
| dc.subject | synovial cells | en |
| dc.subject | anterior cruciate ligament fibroblasts | en |
| dc.subject | cell proliferation | en |
| dc.subject | senescence expression | en |
| dc.title | 幾丁聚醣對前十字韌帶纖維母細胞與滑膜細胞的老化與增生表現之影響 | zh_TW |
| dc.title | The Effect of Chitosan Treatment on the Proliferation and Senescence of Anterior Cruciate Ligament Fibroblasts and Synovial Membrane Derived Cells | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 104-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王至弘(Jyh-Horng Wang),李亦淇(I-Chi Lee),洪智煌(Chih-Huang Hung) | |
| dc.subject.keyword | 前十字韌帶纖維母細胞,滑膜細胞,細胞增生,老化表現,幾丁聚醣, | zh_TW |
| dc.subject.keyword | synovial cells,anterior cruciate ligament fibroblasts,cell proliferation,senescence expression,chitosan, | en |
| dc.relation.page | 54 | |
| dc.identifier.doi | 10.6342/NTU201601048 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2016-07-20 | |
| dc.contributor.author-college | 工學院 | zh_TW |
| dc.contributor.author-dept | 醫學工程學研究所 | zh_TW |
| 顯示於系所單位： | 醫學工程學研究所 | |
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