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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 陳彥榮(Edward Chern) | |
| dc.contributor.author | You-Quan Chen | en |
| dc.contributor.author | 陳宥銓 | zh_TW |
| dc.date.accessioned | 2021-07-10T21:37:02Z | - |
| dc.date.available | 2021-07-10T21:37:02Z | - |
| dc.date.copyright | 2020-08-28 | |
| dc.date.issued | 2020 | |
| dc.date.submitted | 2020-08-18 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76790 | - |
| dc.description.abstract | 間葉幹細胞具有多種可應用於細胞療法的生理特性，在組織再生及免疫疾病等皆具有很高的應用價值。多篇研究表明不同組織來源的間葉幹細胞在生長速率、基因表現、分化以及免疫調節等能力上存在差異表現，可能影響細胞療法的成效。然而這些比較研究受限於培養條件及捐贈者差異對細胞活性的影響，存在結果的不一致。釐清不同組織來源的間葉幹細胞的差異表現將有助於細胞療法的開發。因此，本篇研究統合分析公開核糖核酸測序資料，目的於了解不同組織來源的間葉幹細胞在多篇研究中一致的差異表現。 本篇研究蒐集 114 筆測序資料，並從中篩選出 43 筆分析用資料以及 39 筆驗證用資料。應用分析用資料進行不同組織來源間的差異表現分析，得知不同組織來源的間葉幹細胞間存在大量差異表現基因。經驗證用資料交叉驗證，證實此差異表現基因在所有組別中普遍具有 60 % 以上的預測精密度。以此差異表現基因進行基因本體論分析，得知不同組織來源的間葉幹細胞具體可能在代謝、分化以及調控環境的能力上存在差異。本篇研究也實際取得不同組織來源的間葉幹細胞，發現在驗證的 14 個基因中，11 個基因表現符合差異表現基因的預測。 本篇研究應用統合分析得到不同組織來源的間葉幹細胞的差異表現基因，並透過乾實驗與濕實驗驗證其預測能力。這些差異表現基因可應用於預測特定組織來源的間葉幹細胞擅長的生理功能，進而有助於細胞療法的開發。 | zh_TW |
| dc.description.abstract | Human mesenchymal stem cell (hMSC) has potential in cell therapy targeting tissue regeneration and immune diseases. Studies have shown hMSC derived from different tissues may have different properties, further affecting efficacy of cell therapies. However, specifying differences in hMSCs from comparative studies could be difficult due to their inconsistence in results. Thus, this study conducts a meta-analysis on public RNA-Seq data, aiming to discover consistent differences in previous studies. 43 analyzed data and 39 test data are selected amongst 114 collected RNA-Seq data. Numerous differential expression genes (DEGs) are revealed amongst hMSCs. Precision of these DEGs are cross-validated to be higher than 60 %. Gene ontology analysis shows these DEGs are correlated to metabolism, differentiation, or tissue remodeling. 14 origin-specific marker genes are validated, and 11 correspond to prediction in real samples. These results show that differences in hMSCs can be obtained with RNA-Seq data meta-analysis. Experiments are needed to prove the prediction of functional differences. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-10T21:37:02Z (GMT). No. of bitstreams: 1 U0001-1808202012094200.pdf: 26006061 bytes, checksum: c8276ec0ebe9dbf80fc15c877ac0443f (MD5) Previous issue date: 2020 | en |
| dc.description.tableofcontents | 中文摘要 i 英文摘要 ii 目錄 iii 英文縮寫對照表 vii 第一章 緒論 1 1.1 人類間葉幹細胞 (human mesenchymal stem cell, hMSC) 1 1.2 間葉幹細胞的醫學應用 2 1.3 間葉幹細胞的異質性 (heterogeneity) 3 1.4 培養條件和捐贈者對觀察間葉幹細胞表現的影響 6 1.5 統合分析 (meta-analysis) 7 第二章 研究動機與架構 9 第三章 材料與方法 10 3.1 生物資訊學分析 (bioinformatic analysis) 10 3.1.1 核糖核酸測序公開資料搜索 10 3.1.2 測序資料比對至參考轉錄體 10 3.1.3 轉錄體間的比較分析 11 3.1.4 差異表現基因分析 (differential expression gene analysis) 11 3.1.5 差異表現基因的精密度分析 12 3.1.6 基因本體論分析 (gene ontology analysis) 12 3.1.7 標記基因篩選 (marker gene selection) 13 3.2 人類間葉幹細胞的取得與培養 13 3.2.1 脂肪間葉幹細胞的分離與培養 13 3.2.2 牙髓間葉幹細胞的分離與培養 14 3.2.3 骨髓與臍帶間葉幹細胞的取得與培養 15 3.3 細胞表面標記分析 15 3.4 核糖核酸表現量分析 16 3.4.1 核糖核酸抽取 (RNA extraction) 16 3.4.2 即時定量聚合酶連鎖反應 (Real-Time quantitative PCR) 17 3.5 統計分析 18 第四章 實驗結果 19 4.1 核糖核酸測序公開資料在不同研究計畫間存在明顯的差異 19 4.2 生資分析預測間葉幹細胞間存在大量差異表現基因 20 4.3 統合分析得來之差異表現基因的預測精密度優於單一團隊 21 4.4 基因本體論分析預測間葉幹細胞的功能性差異 21 4.5 生資分析篩選四種組織來源間葉幹細胞的標記基因 22 4.6 即時定量聚合酶連鎖反應驗證生資分析預測的標記基因 23 第五章 綜合討論與未來方向 24 第六章 圖表 28 圖一 主成分分析揭示間葉幹細胞之轉錄體以資料來源團隊分群 28 圖二 相同組織來源的間葉幹細胞在表現浮動基因中以資料來源團隊分群 29 圖三 階層式分群熱度圖揭示間葉幹細胞之轉錄體以資料來源團隊分群 32 圖四 經過篩選的 43 筆分析用資料之轉錄體仍以資料來源團隊分群 33 圖五 不同組織來源的人類間葉幹細胞間存在大量差異表現基因 34 圖六 差異表現基因對驗證用資料表現的預測精密度普遍在 60 % 之上 35 圖七 人類脂肪間葉幹細胞上調之差異表現基因的基因本體論分析 36 圖八 人類骨髓間葉幹細胞上調之差異表現基因的基因本體論分析 39 圖九 人類臍帶間葉幹細胞上調之差異表現基因的基因本體論分析 42 圖十 人類牙髓間葉幹細胞上調之差異表現基因的基因本體論分析 45 圖十一 人類不同組織來源間葉幹細胞的差異表現基因之文氏圖 47 圖十二 人類間葉幹細胞在明視野下呈現紡錘狀的型態 48 圖十三 人類間葉幹細胞表現正確的細胞表面標記分子 49 圖十四 14 個預測的標記基因在人類間葉幹細胞中有 11 個符合預測 50 表一 人類脂肪間葉幹細胞表現浮動基因列表 (前 300) 51 表二 人類骨髓間葉幹細胞表現浮動基因列表 (前 300) 53 表三 人類臍帶間葉幹細胞表現浮動基因列表 (前 300) 55 表四 人類牙髓間葉幹細胞表現浮動基因列表 (前 300) 57 表五 人類脂肪與骨髓間葉幹細胞差異表現基因列表 (部分) 59 表六 人類脂肪與臍帶間葉幹細胞差異表現基因列表 (部分) 60 表七 人類脂肪與牙髓間葉幹細胞差異表現基因列表 (部分) 61 表八 人類骨髓與臍帶間葉幹細胞差異表現基因列表 (部分) 62 表九 人類骨髓與牙髓間葉幹細胞差異表現基因列表 (部分) 63 表十 人類臍帶與牙髓間葉幹細胞差異表現基因列表 (部分) 64 表十一 人類脂肪間葉幹細胞標記基因列表 65 表十二 人類骨髓間葉幹細胞標記基因列表 66 表十三 人類臍帶間葉幹細胞標記基因列表 68 表十四 人類牙髓間葉幹細胞標記基因列表 72 參考文獻 74 附錄一 核糖核酸測序資料清單 i 附錄二 抗體列表 v 附錄三 qPCR 引子列表 vi 附錄四 期刊發表形式 vii | |
| dc.language.iso | zh-TW | |
| dc.subject | 統合分析 | zh_TW |
| dc.subject | 核糖核酸測序 | zh_TW |
| dc.subject | 生物資訊學 | zh_TW |
| dc.subject | 異質性 | zh_TW |
| dc.subject | 人類間葉幹細胞 | zh_TW |
| dc.subject | 組織來源 | zh_TW |
| dc.subject | bioinformatics | en |
| dc.subject | human mesenchymal stem cell | en |
| dc.subject | heterogeneity | en |
| dc.subject | tissue origin | en |
| dc.subject | meta-analysis | en |
| dc.subject | RNA-Seq | en |
| dc.title | 以轉錄體分析探討人類不同組織來源間葉幹細胞之研究 | zh_TW |
| dc.title | Transcriptome Analysis of Human Mesenchymal Stem Cells from Different Tissue Origins | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 108-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 黃兆祺(Eric Hwang),黃楓婷(Feng-Ting Hwang) | |
| dc.subject.keyword | 人類間葉幹細胞,異質性,組織來源,統合分析,核糖核酸測序,生物資訊學, | zh_TW |
| dc.subject.keyword | human mesenchymal stem cell,heterogeneity,tissue origin,meta-analysis,RNA-Seq,bioinformatics, | en |
| dc.relation.page | 81 | |
| dc.identifier.doi | 10.6342/NTU202003945 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2020-08-19 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科技學系 | zh_TW |
| 顯示於系所單位： | 生化科技學系 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| U0001-1808202012094200.pdf 未授權公開取用 | 25.4 MB | Adobe PDF |
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