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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 游佳欣(Jiashing Yu) | |
dc.contributor.author | Chi-Hui Huang | en |
dc.contributor.author | 黃琦惠 | zh_TW |
dc.date.accessioned | 2021-06-16T16:30:32Z | - |
dc.date.available | 2018-01-16 | |
dc.date.copyright | 2013-01-16 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-12-24 | |
dc.identifier.citation | 1. Mohler, E.R., Peripheral arterial disease - Identification and implications. Archives of Internal Medicine, 2003. 163(19): p. 2306-2314.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/63247 | - |
dc.description.abstract | 對於組織的再生和血運重建,植入細胞療法是一個具有極大潛力的治療方式。細胞療法的目標是更換並修復或加強受損組織或器官的功能。然而,有兩個問題是在細胞移植或異種移植前需要解決的主要問題,如此一來才可以達到臨床應用所能接受的標準:控制免疫排斥反應以及控制植入的細胞其增殖的情形。藉由有機或生物衍生之材料,如海藻酸鈉,來包覆細胞的方法,似乎是一個很好的方式來實現上述的目標。
我們利用三維生物培養系統有效的提高了幹細胞的分化能力,且我們藉著利用具有磁懸浮的三維細胞培養系統,即搭配外面塗有仿生物表面的海藻酸鈉基底微載體,來培養初代幹細胞,以使得幹細胞能夠培養維持在三維環境中。 本研究探討了以接枝RGD(精氨酸-甘氨酸-天冬氨酸)之生物性材料實際用於動物模型修復下肢缺血的可能性。在這項實驗中,我們對RGD接枝並混合有高、低兩種分子量的海藻酸鈉微囊球固定脂肪幹細胞hASCs,進行了許多實驗研究。在體外的實驗研究表明,RGD接枝的海藻酸鈉可以成功促進人類脂肪幹細胞(hASCs)的貼附。 基於不同的細胞外基質(ECM)會影響細胞的行為,在接枝RGD的海藻酸鈉固定化細胞方面,顯示出接枝RGD之海藻酸鈉包覆細胞會使得細胞有較高的存活率,整體存活率都高於80%,且有助於促進血管生長因子的表達。我們認為使用成人脂肪幹細胞作為治療下肢缺血患者因病情嚴重而受到影響的病症發展,是個有潛力成為一種成熟且穩定的幹細胞治療方式。 | zh_TW |
dc.description.abstract | Cell-based therapies offer an attractive approach for revascularization and regeneration of tissues. The aim of cell therapy is to replace, repair, or enhance the function of damaged tissues or organs. There are, however, two major drawbacks that need to be resolved before cellular allografts or xenografts can achieve clinical acceptance: control of immune rejection and control of implanted cell proliferation. Cell encapsulation by means of organic or biologically derived matrixes, such as alginate, appears to be an avenue by which these goals may be accomplished.
Three dimensional biology promises to enable efficient differentiation of stem cells. We made use of a magnetically levitating 3D cell culture system, based on alginate microcarriers coated with a biomimetic surface, that supports the growth of primary and stem cells in 3D envirument. The present study was undertaken to explore the possible therapeutic efficacy of RGD (Arg-Gly-Asp) peptide-linked biomaterial in repairing the lower limb ischemia of a rodent model. In this work, hASCs immobilized in RGD-coupled alginate microspheres, with a binary composition of high and low molecular weight alginate, were investigated. The in vitro study of hASCs demonstrated that RGD-coupled alginate promoted cell adherence to the matrix. Since different extra-cellular matrix proteins (ECM) influence the cells behavior, immobilized within RGD-alginate microspheres showed metabolic activity, with an overall viability higher than 80%, growth and increase angiogenic growth factor expression. We believe that the use of adult stem cell therapy in ischemia patients allows for the development of new mature and stable capillaries in patients affected by the severe condition. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T16:30:32Z (GMT). No. of bitstreams: 1 ntu-101-R99524097-1.pdf: 13163615 bytes, checksum: 0e1ab3d152f465e9cad011e94bcdb0d4 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 誌謝 I
摘要 II Abstract III 目錄 IV 圖目錄 VIII 表目錄 XII 第一章 緒論 1 1-1 研究背景 1 第二章 文獻回顧 2 2-1 周邊動脈疾病(peripheral artery disease) 2 2-1-1 周邊動脈疾病之病狀與病徵 3 2-1-2 周邊動脈疾病危險因子 5 2-1-3 周邊動脈疾病篩檢與診斷 7 2-1-4 周邊動脈疾病治療策略 12 2-2 脂肪幹細胞Adipose-derived stem cells (ASCs) 14 2-2-1 脂肪幹細胞來源 14 2-2-2 脂肪幹細胞之萃取 16 2-2-3 脂肪幹細胞之特性 18 2-2-4 脂肪幹細胞治療周邊動脈疾病之潛能 20 2-2-5 脂肪幹細胞促進血管新生之機制 20 2-2-6 血管內皮生長因子(VEGF) 23 2-3 組織工程(tissue engineering) 24 2-3-1 細胞外間質(extracellular matrix) 25 2-3-2 二維細胞培養 27 2-3-3 三維細胞培養 29 2-3-4 三維生物反應器 30 2-3-5 三維生物反應器種類 31 2-3-6 微載體(microcarrier) 33 2-4 細胞固定化 35 2-4-1 膠體包埋法(Matrix entrapment) 36 2-4-2 交聯法(Cross-linking binding) 36 2-4-3 擔體鍵結法(Binding to carriers) 36 2-4-4 薄膜包覆法(Membrane enclosure) 36 2-5 海藻膠(Alginate) 37 2-5-1 海藻膠之凝膠性質 38 2-6生醫材料與細胞貼附 40 2-6-1細胞活化因子修飾生醫材料 42 第三章 材料與方法 44 3-1 實驗設備 44 3-2 實驗藥品 46 3-3 實驗步驟 50 3-3-1 脂肪幹細胞的萃取與培養 50 3-3-1-1細胞萃取步驟 Stromal Vascular Fraction(SVF) 50 3-3-1-2細胞繼代培養(Subculture) 53 3-3-1-3細胞冷凍保存: 54 3-3-2 微載體三維培養系統 54 3-3-2-1 脂肪幹細胞貼附 57 3-3-2-2 微載體細胞貼附增生測試(MTT assay) 58 3-3-2-3 脂肪幹細胞貼附蛋白質定量分析(BCA assay) 60 3-3-2-4 脂肪幹細胞三維培養 61 3-3-2-5 脂肪幹細胞三維培養計數 67 3-3-2-5 脂肪幹細胞三維培養螢光染色 69 3-3-3 脂肪幹細胞分化 70 3-3-3-1 分化培養基底配製 70 3-3-3-2 脂肪分化誘導培養基配製 71 3-3-3-3 硬骨分化誘導培養基配製 72 3-3-3-4 油紅染劑配製 72 3-3-3-5 茜素紅染劑配製 72 3-3-3-6 鹼性磷酸酶染劑配製 73 3-3-3-7 脂肪細胞染色 73 3-3-3-8 硬骨細胞染色 74 3-3-3-9 脂肪分化之三酸甘油脂定量分析 76 3-3-3-10 硬骨分化之鈣沉積量定量分析 77 3-3-4 微囊固定化脂肪幹細胞 78 3-3-4-1 海藻酸鈉(sodium alginate)改質 79 3-3-4-2 海藻酸鈉細胞貼附增生測試(MTT assay) 84 3-3-4-3 微囊包覆機製作海藻酸鈉微球 85 3-3-4-3 海藻酸鈉微囊球粒徑測試 89 3-3-4-4 海藻酸鈉微囊球細胞計數 89 3-3-4-5 海藻酸鈉微囊球細胞螢光染色 89 3-3-4-6 海藻酸鈉微囊球細胞活性測試(Live/Dead Viability) 90 3-3-4-7 ELISA免疫分析法(Enzyme-linked immunosorbent assay) 91 3-3-4-8 蛋白質電泳及西方墨點法(SDS-PAGE and Western blot) 92 3-3-4-9 反轉錄聚合酵素連鎖反應(RT-PCR) 92 3-3-4-10掃瞄式電子顯微鏡(SEM)影像 93 第四章 結果與討論 94 4-1脂肪幹細胞的萃取與培養 94 4-1-1 脂肪幹細胞萃取 Stromal Vascular Fraction(SVF) 94 4-2 微載體三維培養系統 96 4-2-1 微載體細胞貼附增生測試 96 4-2-2 脂肪幹細胞三維培養 102 4-2-3 脂肪幹細胞三維培養計數 112 4-3 三維v.s二維脂肪幹細胞(ASCs)分化能力測定 114 4-3-1 ASCs之脂肪細胞分化能力測定 114 4-3-2 ASCs之骨細胞分化能力測定 117 4-4 微囊固定化脂肪幹細胞 121 4-4-1 海藻酸鈉改質測定 121 4-4-2海藻酸鈉細胞貼附增生測試 124 4-4-3 海藻酸鈉微囊球粒徑測試 127 4-4-4 海藻酸鈉微囊球計數與細胞活性 129 4-4-5反轉錄聚合酵素連鎖反應(RT-PCR) 134 第五章 結論 135 第五章 未來展望 136 參考文獻 137 | |
dc.language.iso | zh-TW | |
dc.title | RGD改質海藻酸鈉微球包覆三維培養人類脂肪幹細胞 | zh_TW |
dc.title | Immobilization of 3D culture human adipose-derived stem cells within RGD modified alginate microspheres | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蔡偉博(Wei-Bor Tsai),廖英志(Ying-Chih Liao),陳賢燁(Hsien-Yeh Chen) | |
dc.subject.keyword | 三維細胞培養,微載體,RGD修飾,海藻酸鈉,下肢缺血疾病, | zh_TW |
dc.subject.keyword | 3D cell culture,microcarrier,RGD-modified,Alginate,PAD, | en |
dc.relation.page | 149 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-12-25 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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