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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 何弘能(Hong-Nerng Ho) | |
| dc.contributor.author | Chen-Wei Lan | en |
| dc.contributor.author | 藍晨瑋 | zh_TW |
| dc.date.accessioned | 2021-06-16T08:11:26Z | - |
| dc.date.available | 2019-10-15 | |
| dc.date.copyright | 2014-10-15 | |
| dc.date.issued | 2014 | |
| dc.date.submitted | 2014-02-19 | |
| dc.identifier.citation | (2004)'Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome.' Fertil Steril81(1): 19-25.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/58325 | - |
| dc.description.abstract | 卵巢顆粒細胞在卵子成熟生長過程中扮演相當重要的角色。研究指出顆粒細胞失去功能易造成卵巢功能異常，例如多囊性卵巢症候群。
人類胚胎幹細胞具有無限的自我更新能力，並能分化成各種細胞類型。經由多種步驟，利用不同之生長因子及合成蛋白刺激人類胚胎幹細胞之分化，以基因表達來檢測從胚胎幹細胞分化經原條內胚層及中間板中胚層，最後分化到具有功能性表達之顆粒細胞特異性轉錄因子FOXL2、雌激素合成酶CYP19A1、抗穆勒氏管激素AMH、第二型AMH的受體AMHR2、卵泡刺激素受體FSHR及具有分泌AMH和轉化睾丸激素形成雌激素之能力。這項研究所建立的系統將提供一個模式探討卵巢顆粒細胞發育。 為了更瞭解多囊性卵巢症候群在卵巢顆粒細胞上所造成的病因，我們利用基因微陣列技術比較6名健康女性及6名多囊性卵巢症候群患者之顆粒細胞間基因差異。研究數據分析顯示，總共有243個基因在正常對照組與多囊性卵巢症患者之顆粒細胞間具有顯著差異。這些有差異的基因主要與生殖系統之發育與功能有關。比較分析顯示也涉及MAPK/ERK訊號傳遞路徑的調控。利用西方點墨法分析更證實並驗證在多囊性卵巢症患者之顆粒細胞MAP3K4及phospho-ERK1/2活性明顯降低。 這項研究找到了許多新的基因有可能在卵巢顆粒細胞的功能上扮演重要角色，並可能是影響多囊性卵巢症患者可能的遺傳病因。研究中也發現多囊性卵巢症患者之顆粒細胞與MAPK/ERK訊號傳遞路徑的調控有關。這個分子機制將進一步幫助我們了解發病機制並開發新療法。 | zh_TW |
| dc.description.abstract | Ovarian granulosa cells are important for the development of oocytes. Recent studies have suggested that the dysfunction of granulosa cells may lead to disorders of ovarian function, such as polycystic ovary syndrome (PCOS).
Human embryonic stem cells (hESCs) have an unlimited capacity and can differentiate into all cell types. Through multi-step approaches comprising in vitro treatments with cocktails of growth factors, gene expression analyse the progress of hESCs to primitive streak–mesendoderm, intermediate plate mesoderm, and finally to functional granulosa-like cells that expressed the granulosa cell-specific forkhead transcription factor FOXL2, CYP19A1, anti-Müllerian hormone, the type 2 AMH receptor, and the follicle stimulating hormone receptor. The granulosa cells are capable of producing AMH and aromatizing testosterone to estradiol. This developmental system will provide a model to elucidate granulosa cell development. In order to understand PCOS disease on the granulosa cells, we use cDNA microarray technology to compare differential gene expressions in granulosa cells: including 6 women with normal ovulatory and 6 with PCOS. A total of 243 genes were showed to have differential expressions in the PCOS granulosa cells when compared with normal. These differentially expressed genes are involved in reproductive system development and involved in the mitogen-activated protein kinase/ extracellular regulated kinase (MAPK/ERK) signaling pathways. Western blot analyses confirmed that MAP3K4 and phospho-ERK1/2 was decreased in PCOS granulosa cells. The molecular mechanisms will help us to understand the pathogenesis for PCOS disease. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T08:11:26Z (GMT). No. of bitstreams: 1 ntu-103-D98421003-1.pdf: 2874904 bytes, checksum: 3afcbbbeb63df7de37fbcd900ce5f44e (MD5) Previous issue date: 2014 | en |
| dc.description.tableofcontents | 目錄 (Index)...............................................1
圖目錄 (Figure index)......................................3 表目錄 (Table index).......................................5 英文縮寫表 (Abbreviations).................................6 摘要 (Abstract):中文摘要...................................8 英文摘要...................................9 第一章 緒論 (Introduction)................................10 第一節 卵巢顆粒細胞在卵泡形成中扮演的重要性及發育生物學...10 第二節 多囊性卵巢症候群 (polycystic ovary syndrome).......13 第三節 卵巢顆粒細胞的研究及與多囊性卵巢症候群之關係.......14 第四節 人類多潛能幹細胞的研究及應用.......................15 第五節 人類多潛能幹細胞目前應用與顆粒細胞間之研究展望.....20 第六節 研究目的與假說.....................................21 第二章 研究材料及方法 (Methods and Materials).............23 第一部份 人類胚胎幹細胞分化成具功能性之卵巢顆粒細胞 第一節 細胞培養及卵巢顆粒細胞之分離 (cell culture and isolation)................................................23 第二節 人類胚胎幹細胞經由中胚層分化成顆粒細胞之方法.......25 第三節 免疫螢光染色 (immunofluorescence staining).........26 第四節 RNA萃取及即時定量聚合酶連鎖反應 (real-time PCR)....27 第五節 蛋白質萃取及西方點墨法 (Western blot analyses).....28 第六節 流式細胞儀分析 (flow cytometric analysis)..........29 第七節 芳香環化酶活性 (aromatase activity) 及雌二醇之檢測試........................................................30 第八節 抗穆氏管荷爾蒙 (AMH) 之檢測........................31 第九節 統計分析 (statistical analysis)....................32 第二部份 多囊性卵巢症候群之基因差異性及機轉研究 第一節 病人的篩選 (patient selection).....................33 第二節 卵巢顆粒細胞的分離及RNA之萃取......................33 第三節 DNA微陣列雜合技術 (DNA microarray hybridization)...34 第四節 蛋白質萃取及西方點墨法 (Western blot analyses).....35 第五節 微陣列資料處理與統計分析 (IPA & statistical analysis).................................................36 第三章 研究結果 (Results).................................37 第一部份 人類胚胎幹細胞分化成具功能性之卵巢顆粒細胞 第一節 人類胚胎幹細胞經由類胚體形成方式往中胚層分化.......37 第二節 人類胚胎幹細胞衍生之中胚層細胞分化成卵巢顆粒細胞...38 第三節 睾固酮經由人類胚胎幹細胞衍生的類顆粒細胞轉化成雌二醇........................................................39 第四節 人類胚胎幹細胞衍生的類顆粒細胞產生抗穆氏管荷爾蒙 (AMH).....................................................40 第二部份 多囊性卵巢症候群之基因差異性及機轉研究 第一節 正常對照組與多囊性卵巢症候群之臨床診斷與特徵.......41 第二節 正常對照組與多囊性卵巢症候群之基因差異表達譜.......41 第三節 多囊性卵巢症基因差異及其相關功能、網路和訊息傳導路徑........................................................41 第四節 利用西方點墨法驗證微陣列資料之結果.................42 第四章 討論 (Discussion)..................................44 第五章 未來展望 (Perspectives) 及應用 (Application).......49 第六章 論文英文簡述 (Summary in English)..................53 第七章 參考文獻 (Reference)...............................81 第八章 圖表 (Figures and Tables)..........................94 第九章 附錄 (Appendix)...................................122 | |
| dc.language.iso | zh-TW | |
| dc.subject | 人類胚胎幹細胞 | zh_TW |
| dc.subject | 卵巢顆粒細胞 | zh_TW |
| dc.subject | 濾泡生成 | zh_TW |
| dc.subject | 抗穆氏管荷爾蒙 | zh_TW |
| dc.subject | 多囊性卵巢症候群 | zh_TW |
| dc.subject | MAPK/ERK訊號傳遞路徑 | zh_TW |
| dc.subject | MAPK/ERK signal pathway | en |
| dc.subject | polycystic ovary syndrome | en |
| dc.subject | human embryonic stem cells | en |
| dc.subject | granulosa cells | en |
| dc.subject | folliculogenesis | en |
| dc.subject | anti-mullerian hormone | en |
| dc.title | 將人類胚胎幹細胞分化為卵巢顆粒細胞：可應用於多囊性卵巢症之治療及基因機轉之探討 | zh_TW |
| dc.title | Differentiation of human embryonic stem cells into granulosa cells: potential application in polycystic ovary syndrome and investigation of genetic mechanisms | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.coadvisor | 陳信孚(Hsin-Fu Chen) | |
| dc.contributor.oralexamcommittee | 高嘉宏,林泰元,沈家寧,黃彥華 | |
| dc.subject.keyword | 人類胚胎幹細胞,卵巢顆粒細胞,濾泡生成,抗穆氏管荷爾蒙,多囊性卵巢症候群,MAPK/ERK訊號傳遞路徑, | zh_TW |
| dc.subject.keyword | human embryonic stem cells,granulosa cells,folliculogenesis,anti-mullerian hormone,polycystic ovary syndrome,MAPK/ERK signal pathway, | en |
| dc.relation.page | 122 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2014-02-19 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
| 顯示於系所單位： | 臨床醫學研究所 | |
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