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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳玉怜(Yuh-Lien Chen) | |
| dc.contributor.author | Shu-Huei Wang | en |
| dc.contributor.author | 王淑慧 | zh_TW |
| dc.date.accessioned | 2021-06-15T01:51:41Z | - |
| dc.date.available | 2009-09-15 | |
| dc.date.copyright | 2009-09-15 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-03 | |
| dc.identifier.citation | Alobaid N, Alnaeb ME, Sales KM, Seifalian AM, Mikhailidis DP, Hamilton G. Endothelial progenitor cells and their potential clinical applications in peripheral arterial disease. Endothelium. 2005; 12:243-250.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43357 | - |
| dc.description.abstract | 在以細胞植入治療心血管疾病方面，欲由骨髓及周邊血液中分離出具有潛能的內皮前驅細胞，結果發現骨髓及周邊血液中的內皮前驅細胞不論在數量或品質上都受到相當大的限制。本篇研究之主要目的為檢測可否從臍帶間質(Wharton’s jelly in human umbilical cord, WJC)中分離、培養並分化出內皮前驅細胞，及探討其是否可影響內皮受損後的內膜增生。首先由臍帶間質分離出的間質細胞(mesenchymal cells, MCs)，培養於適合內皮細胞生長的培養液(EGM-2)中可以分化成內皮前驅細胞(late outgrowth endothelial cells) (WJC-OECs)，其具有吞噬乙醯化的低密度脂蛋白(acetylated-LDL)之能力，並且表現出內皮細胞特有的蛋白。利用彈性線圈將老鼠股動脈進行去內皮手術後，再將這些已分化的人類內皮前驅細胞經由尾靜脈注射植入老鼠體內，結果發現，這些植入的細胞能夠快速到達內皮受損區，進行血管內皮修補(re-endothelialization)的作用。在去內皮手術後及細胞植入4周後，相較植入saline、MCs及臍帶血衍生之內皮前驅細胞(cord blood-OEC, CB-OECs)之組別，發現植入WJC-OECs之組別中，血管內膜層與中膜層之比例有顯著的降低(2周組，saline: 1.48±0.14, MC: 1.32±0.18, WJC-OEC: 0.95±0.05, CB-OEC: 1.32±0.05; 4周組，分別為saline: 1.81±0.25, MC: 1.68±0.09, WJC-OEC: 01.01±0.07, CB-OEC: 1.39±0.14)，而且在內皮受損區，經由組織免疫染色法觀察有大量的色素性上皮細胞衍生因子(pigment epithelium-derived factor, PEDF)表現，相同的結果也可在細胞培養時觀察到；進一步發現WJC-OECs的培養液會抑制人類主動脈平滑肌細胞移行及增生；而藉由加入PEDF抗體或轉染PEDF siRNA來減弱WJC-OECs之PEDF的表現後，顯著抑制其影響。本篇報告是第一篇陳述臍帶間質細胞具有分化成內皮前驅細胞的潛能，而且對於血管內皮細胞受損後的再修補扮演著相當重要的角色，可維持血管內皮的完整，達到抑制內膜增厚的作用。綜合本研究的結果顯示，在血管受損相關疾病的臨床治療，WJC-OEC可視為細胞治療法中的另一個新契機。 | zh_TW |
| dc.description.abstract | The number of endothelial progenitor cells (EPCs) that can be obtained from adult bone marrow and peripheral blood to treat cardiovascular diseases is limited. The goal was to examine the endothelial potential of Wharton’s jelly in human umbilical cord (WJC)-derived stem cells and evaluate their potential to affect neointimal formation after vascular injury. Mesenchymal cells (MCs) were isolated from WJC and cultured in endothelial growth medium. Differentiation into late outgrowth endothelial cells (WJC-OECs) was demonstrated by incorporation of acetylated low-density lipoprotein and expression of the endothelial-specific markers. Transplantation of these cells into wire-injured femoral arteries in mice led to rapid re-endothelialization. At 4 weeks after injury, the neointima/media area ratio was reduced and strong expression of pigment epithelium-derived factor (PEDF) compared to saline- or MC- or cord blood-OEC- treated mice. WJC-OECs-conditioned medium has an extremely strong capacity to inhibit the migration and proliferation of smooth muscle cells. The effects were inhibited by neutralizing antibody for PEDF and by siRNA silencing of PEDF. We firstly demonstrated the presence of a cell population within WJC that has the potential to differentiate into OECs. Transplantation of WJC-OECs may play a crucial role in reestablishing endothelial integrity in injured vessels, thereby inhibiting neointimal hyperplasia. These findings have implications for a novel and practical cell-based therapy for vascular diseases. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T01:51:41Z (GMT). No. of bitstreams: 1 ntu-98-D94446004-1.pdf: 9570725 bytes, checksum: b4f08387f637112a29ac595e8ce3d70a (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 英文縮寫與全名對照表------------------------------------------------------------------------I
英文摘要-----------------------------------------------------------------------------------------------III 中文摘要------------------------------------------------------------------------------------------------V 文獻回顧-------------------------------------------------------------------------------------------------1 一 引言-------------------------------------------------------------------------------------------------1 二 動脈粥狀硬化及血管(再)狹窄之成因-----------------------------------------------------1 三 內皮細胞及平滑肌細胞與心血管疾病之關係------------------------------------------3 四 前驅細胞之來源種類及功能應用----------------------------------------------------------4 五 臍帶間質組織之特性及應用----------------------------------------------------------------6 六 內皮前驅細胞之定義、鑑定、分離培養及表現型-----------------------------------7 七 PEDF與動脈粥狀硬化及血管再狹窄之關聯------------------------------------------8 研究動機及實驗設計-------------------------------------------------------------------------------10 儀器設備-----------------------------------------------------------------------------------------------11 實驗材料與試劑--------------------------------------------------------------------------------------11 實驗用溶液配方--------------------------------------------------------------------------------------15 材料與方法--------------------------------------------------------------------------------------------17 一 細胞初級培養---------------------------------------------------------------------------17 人類臍帶間質細胞及人類臍帶間質內皮前驅細胞之培養-----------------17 人類臍帶臍靜脈內皮細胞之培養-------------------------------------------------17 人類臍帶血內皮前驅細胞之培養-------------------------------------------------18 人類主動脈平滑肌細胞之培養-----------------------------------------------------18 二 內皮細胞功能之定性------------------------------------------------------------------18 DiI-acetylated LDL uptake -------------------------------------------------------18 Matrigel tube-formation-----------------------------------------------------------19 三 前驅細胞潛力之比較------------------------------------------------------------------19 Colony forming assay----------------------------------------------------------------19 Replicative capacity------------------------------------------------------------------19 四 PEDF siRNA----------------------------------------------------------------------------19 五 酵素免疫分析法------------------------------------------------------------------------20 六 動物股動脈去內皮傷害模式及內皮前驅細胞植入--------------------------20 七 免疫組織化學染色---------------------------------------------------------------------21 八 西方墨點法-------------------------------------------------------------------------------21 九 細胞免疫螢光染色---------------------------------------------------------------------22 十 流式細胞儀分析-----------------------------------------------------------------------23 十一 細胞傷口癒合實驗---------------------------------------------------------------------23 十二 細胞穿膜試驗---------------------------------------------------------------------------24 十三 BrdU incorporation staining--------------------------------------------------------24 十四 反轉錄-聚合酶連鎖反應--------------------------------------------------------------24 十五 數據統計分析----------------------------------------------------------------------------25 結果------------------------------------------------------------------------------------------------------26 壹 臍帶間質細胞具有分化成內皮前驅細胞之能力-----------------------------------26 貳 內皮前驅細胞抑制因內皮細胞損傷所引致血管內膜增生----------------------27 參 內皮前驅細胞所釋出PEDF具有抑制內膜增生的作用--------------------------28 肆 分化後內皮前驅細胞釋出PEDF具有抑制平滑肌細胞移行效用-------------28 伍 分化後內皮前驅細胞釋出PEDF具有抑制平滑肌細胞增生作用-------------29 陸 分化後內皮前驅細胞釋出的PEDF對於內皮細胞沒有顯著抑制移行及增生之作用-------------------------------------------------------------------------------------------30 討論------------------------------------------------------------------------------------------------------32 結論與未來展望--------------------------------------------------------------------------------------36 參考文獻-----------------------------------------------------------------------------------------------37 附圖------------------------------------------------------------------------------------------------------51 圖一 初級培養間質細胞(MC)之型態及分化----------------------------------------------51 圖二 臍帶內皮前驅細(WJC-OEC)之特徵--------------------------------------------------52 圖三 初級培養臍帶血內皮前驅細(CB-OEC)之型態及分化---------------------------53 圖四 臍帶血內皮前驅細(CB-OEC)之定性-------------------------------------------------54 圖五 內皮前驅細胞內CD133之表現情形---------------------------------------------------55 圖六 股動脈在去內皮(denudation)手術後立即植入細胞，在4小時後，植 入的細胞在受傷對側正常血管切片上的表現情形--------------------------56 圖六 股動脈在去內皮(denudation)手術後立即植入細胞，在24小時後，植入的細胞在受傷對側正常血管切片上的表現情形-----------------------57 圖七 股動脈在去內皮(denudation)手術後立即植入細胞，在4及24小時後，植入的細胞皆未出現在受傷對側正常血管切片上----------------------58 圖八 植入的WJC-OEC能有效減少股動脈因內皮細胞損傷所引致的內膜增厚 ----------------------------------------------------------------------------------------------------59 圖九 各組細胞PEDF之表現量-----------------------------------------------------------------61 圖十 WJC-OEC-CM及PEDF能有效抑制HASMC之移行--------------------------62 圖十一 WJC-OEC-CM及PEDF能有效抑制HASMC之增生-----------------------64 圖十二 WJC-OEC-CM及PEDF對HUVEC移行之效應------------------------------65 圖十三 WJC-OEC-CM及PEDF對HUVEC增生之效應------------------------------67 | |
| dc.language.iso | zh-TW | |
| dc.title | 臍帶間質組織衍生之內皮前驅細胞具有抑制內皮受損後內膜增生之作用: 藉由色素性上皮細胞衍生因子參與抑制作用 | zh_TW |
| dc.title | Late outgrowth endothelial cells derived from Wharton’s jelly in human umbilical cord reduce neointimal formation after vascular injury: involvement of pigment epithelium-derived factor | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 王淑美(Shue-Mei Wang),吳華林(Hua-Lin Wu),裘正健(Jeng-Jiann Chiu),王懷詩(Hwai-Shi Wang),陳永祥(Yung-Hsiang Chen) | |
| dc.subject.keyword | 內皮前驅細胞,再狹窄,脊髓間質組織,細胞治療,色素性上皮細胞衍生因子, | zh_TW |
| dc.subject.keyword | endothelial progenitor cells,restenosis,Wharton’s jelly,cell therapy,pigment epithelium-derived factor (PEDF), | en |
| dc.relation.page | 67 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2009-07-03 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 解剖學暨生物細胞學研究所 | zh_TW |
| 顯示於系所單位： | 解剖學暨細胞生物學科所 | |
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