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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊台鴻(Tai-Horng Young) | |
| dc.contributor.author | Jui-Nan Lu | en |
| dc.contributor.author | 陸瑞男 | zh_TW |
| dc.date.accessioned | 2021-06-12T18:31:05Z | - |
| dc.date.available | 2011-08-22 | |
| dc.date.copyright | 2011-08-22 | |
| dc.date.issued | 2011 | |
| dc.date.submitted | 2011-08-08 | |
| dc.identifier.citation | Background
1. Liu HC, Lee IC, Wang JH, Yang SH, Young TH. Preparation of PLLA membranes with different morphologies for culture of MG-63 Cells. Biomaterials 2004 Aug;25(18):4047-4056. 2. Hung CH, Lin YL, Young TH. The effect of chitosan and PVDF substrates on the behavior of embryonic rat cerebral cortical stem cells. Biomaterials 2006 Sep;27(25):4461-4469. 3. Hoshiba T, Cho CS, Murakawa A, Okahata Y, Akaike T. The effect of natural extracellular matrix deposited on synthetic polymers on cultured primary hepatocytes. Biomaterials 2006 Sep;27(26):4519-4528. 4. Verma P, Verma V, Ray P, Ray AR. Formation and characterization of three dimensional human hepatocyte cell line spheroids on chitosan matrix for in vitro tissue engineering applications. In vitro cellular & developmental biology 2007 Nov-Dec;43(10):328-337. 5. Sanchez A, Alvarez AM, Pagan R, Roncero C, Vilaro S, Benito M, et al. Fibronectin regulates morphology, cell organization and gene expression of rat fetal hepatocytes in primary culture. Journal of hepatology 2000 Feb;32(2):242-250. 6. Wang CC, Lu JN, Young TH. The alteration of cell membrane charge after cultured on polymer membranes. Biomaterials 2007 Feb;28(4):625-631. Part I 1. Katsura N, Ikai I, Mitaka T, Shiotani T, Yamanokuchi S, Sugimoto S, et al. Long-term culture of primary human hepatocytes with preservation of proliferative capacity and differentiated functions. The Journal of surgical research 2002;106(1):115-123. 2. Zhao P, Kunze KL, Lee CA. Evaluation of time-dependent inactivation of CYP3A in cryopreserved human hepatocytes. Drug metabolism and disposition: the biological fate of chemicals 2005;33(6):853-861. 3. Hewitt NJ, Buhring KU, Dasenbrock J, Haunschild J, Ladstetter B, Utesch D. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/27974 | - |
| dc.description.abstract | 在此研究中，我們利用高分子基材培養冷凍四年的人類肝臟細胞，結果顯示肝臟細胞在冷凍四年後在聚乙烯醇上仍然可以保有肝臟細胞的功能及活性，因此我們認為聚乙烯醇是一個未來能夠應用在培養冷凍肝臟細胞的有潛力之高分基材。在第二部分我們以同時含有幾丁聚醣及組織培養的聚苯乙烯之複合材料培養環境去培養人類肝臟的細胞株HepG2，從實驗中我們可以知道決定一個細胞的行為表現主要的因素是來自於細胞基底下面的基材，景館一個環境中還有兩種行為表現不同的細胞存在，彼此並不會相互影響而改變細胞在基材上的特性。在第三部分，我們利用胰島素的添加去改變細胞在幾丁聚醣上的行為表現，從實驗中我們知道利用胰島素的添加可以明顯的提升肝臟細胞在幾丁聚醣上的貼付能力與增生的能力，此外我們同樣的利用抑制PI-3 kinase路徑的藥劑來終止胰島素對細胞行為的影響。在最後的部分，我們利用細胞電泳來偵測神經幹細胞初期的分化。此外我們發現神經幹細胞初期的分化會伴隨著表面電位的提升。 | zh_TW |
| dc.description.abstract | The purpose of the first part was to evaluate the behaviors of long-term (more than 4 years) cryopreserved human hepatocytes on biomaterials. Human hepatocytes could be cryopreserved for more than 4 years without losing liver-specific function and PVA was proposed to serve as an appropriate and promising substrate for the use in culturing long-term cryopreserved hepatocytes by maintaining high cell attachment and of high level of liver-specific function. In the second part, cells were cultured on a multi-biomaterials substrate (C/T substrate) both composed of chitosan and TCPS (tissue culture polystyrene) for 8 days. Cells expressed the behaviors based on the substrate under the cells, and none of cell was found that migrated from one substrate to the other substrate. And in the third part, insulin was added into the CT substrate to control the cell behaviors on the chitosan. Insulin greatly promted cell adhesion and enhanced the proliferation on chitosan. The insulin induced cellular behaviors can be removed by blocking the PI-3 kinase pathway, and cell behaviors restored to the chitosan-induced behaviors. And in the final part, macroelectrophoresis was used to detected the differentiation of neural stem/progenitor cell (NSPC). We found that the surface charge of NSPC was up-regulated during the early period of differentialtion. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-12T18:31:05Z (GMT). No. of bitstreams: 1 ntu-100-D93548005-1.pdf: 3275459 bytes, checksum: e2608fe169b5a34beb18b27afca18280 (MD5) Previous issue date: 2011 | en |
| dc.description.tableofcontents | 致謝 I
ABSTRACT II 摘要 III BACKGROUND 1 REFERENCES 3 PART I THE SUITABLE BIOMATERIALS FOR CULTURING LONG TERM CRYOPRESERVED HUMAN PRIMARY HEPATOCYES 4 INTRODUCTION 5 MATERIALS AND METHODS 7 Preparation of culturing substrates 7 Isolation of human hepatocytes 8 Cryopreservation and thawing of human hepatocytes 8 Culture of post-thaw human hepatocytes 10 Cell attachment assay 10 Cell viability and function assay 11 Cell morphology assay 12 RESULTS 13 Cell attachment 13 Cell viability 15 Albumin secretion 16 Urea synthesis 16 DISCUSSION 17 REFERENCES 22 TABLE AND FIGURES 25 PART II THE CELL BEHAVIORS OF HEPG2 ON THE CHITOSAN/TCPS COMBINDED SUBSTRATE 31 INTRODUCTION 32 MATERIALS AND METHODS 34 Preparation chitosan and C/T substrates 34 Cell culture and assays for cell morphology 35 Assessment of cell proliferation and death 35 SDS-PAGE/Western Blotting 36 The adsorbed fibronectin on the chitosan and TCPS substrates 37 RESULTS 38 Cell morphology 38 Cell migration 39 Cell proliferation and death 40 Cell function 41 Synthesis and adsorption of fibronectin 41 DISCUSSION 42 REFERENCE 48 FIGURES 50 PART III THE EFFECTIVENESS OF INSULIN ON THE CELLULAR BEHAVIORS OF HEPG2 ON THE CHITOSAN 58 INTRODUCTION 59 MATERIALS AND METHODS 60 Preparation chitosan and C/T substrates 60 Cell culture and assays for cell morphology 61 Assessment of cell proliferation and death 61 SDS-PAGE/Western Blotting 62 RESULTS 63 The effects of the insulin on the behaviors of HepG2 on the chitosan 63 Cell morphology 63 Cell proliferation and liver specific function 64 The effects of the insulin on the behaviors of HepG2 on the chitosan/TCPS (C/T) combined substrates. 66 Cell morphology and the behaviors of cells at the boundary of chitosan and TCPS in C/T substrate 66 Cell growth and liver specific function 67 The behaviors of insulin-treated HepG2 after insulin were removed from culture system. 69 The alteration of insulin on the behaviors of HepG2 can be related to PI-3 kinase pathway. 69 DISCUSSION 71 REFERENCE 75 FIGURES 78 PART IV THE APPLICATION OF MACROELECTROPHOSESIS ON THE EARLY DETECTION OF THE DIFFERENTIATED STEM CELLS 86 INTRODCTION 87 MATERIALS AND METHODS 88 Materials 88 Preparation of polymer substrates 89 Culture of neural stem/progenitor cells (NSPCs) 89 Immunocytochemistry 91 Assessment of cell electrophoretic mobility 92 Quantification of migrated out NSPCs from neurospheres 93 RESULTS 94 Electrophoretic mobility of NSPCs on PVDF and TCPS 94 The effect of serum on electrophoretic mobility of NSPCs 96 The effect of PDL on electrophoretic mobility of NSPCs 97 The relationship between the electrophoretic mobility and differentiation of NSPCs 98 The regulation of NSPC differentiation by controlling cell electrophoretic mobility 100 DISCUSSION 102 NSPC differentiation accompanied by elevation of cellular electrophoretic mobility. 102 The energy barrier of NSPC differentiation. 104 The role of high activation energy in maintaining the stability of NSPCs. 105 To regulate NSPC differentiation by controlling the cellular electrophoretic mobility. 106 REFERENCES 109 FIGURES 111 | |
| dc.language.iso | en | |
| dc.subject | 細胞電泳 | zh_TW |
| dc.subject | 人類肝細胞 | zh_TW |
| dc.subject | 冷凍保存 | zh_TW |
| dc.subject | HepG2細胞株 | zh_TW |
| dc.subject | 幾丁聚醣 | zh_TW |
| dc.subject | hepatocyte | en |
| dc.subject | macroelectrophoresis | en |
| dc.subject | chitosan | en |
| dc.subject | HepG2 | en |
| dc.subject | cryopreservation | en |
| dc.title | 人類肝臟細胞在高分子基材上之行為表現及細胞電永在組織工程上的應用 | zh_TW |
| dc.title | The study pf behaviors of hepatocyte on different biomaterials and the application of macroelectrophoresis on tissue engineering | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 99-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 鄭廖平(Liao-Ping Cheng),林達鎔(Dar-Jong Lin),林宏殷(Hung-Yin Lin),李玫樺(Mei-Hwa Lee),胡威文(Wei-Wen Hu),洪智煌 | |
| dc.subject.keyword | 人類肝細胞,冷凍保存,HepG2細胞株,幾丁聚醣,細胞電泳, | zh_TW |
| dc.subject.keyword | hepatocyte,cryopreservation,HepG2,chitosan,macroelectrophoresis, | en |
| dc.relation.page | 126 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2011-08-08 | |
| 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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| ntu-100-1.pdf 未授權公開取用 | 3.2 MB | Adobe PDF |
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