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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳敏慧 | |
dc.contributor.author | Yuan-Yu Tu | en |
dc.contributor.author | 杜元佑 | zh_TW |
dc.date.accessioned | 2021-06-07T18:09:24Z | - |
dc.date.copyright | 2012-09-17 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-07-11 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16314 | - |
dc.description.abstract | 實驗目的: 人類乳牙牙髓幹細胞(stem cells from human exfoliated deciduous teeth,SHED)和人類牙根尖幹細胞(stem cells from apical papilla,SCAP)是屬於多效性的幹細胞(Multipotent stem cell),兩種細胞經過誘導都有分化成神經細胞的能力。我們的假設是乳牙牙髓幹細胞與人類牙根尖幹細胞具有相同的神經分化能力。本實驗的目的是希望經過誘導神經分化(Neurodifferentiation)作用後比較兩種細胞神經分化的能力。
實驗方法:將兩種細胞培養誘導神經分化培養液(Neural induction medium)中一天、三天、七天、十四天、二十一天,以MTT計算活細胞的量,培養零天、一天、三天、七天、十四天、二十一天用光學顯微鏡直接觀察細胞外型變化以及反轉錄聚合酶反應(RT-PCR)和免疫螢光染色(Immuoflurorescence)來比較兩者神經分化的能力。 實驗結果:在光學顯微鏡觀察下兩種細胞在三天後就會有部分細胞變成類似神經細胞外型的改變,而在MTT的實驗中可以發現細胞在接受神經分化之培養液作用後具有細胞增生力會降低,利用反轉錄聚合酶反應和免疫螢光染色實驗發現兩種細胞具有分化成神經細胞的能力。而經過誘導後的人類牙根尖幹細胞神經纖維(Neurofilament)基因之表現上比經過誘導後乳牙牙髓幹細胞高,顯示人類牙根尖幹細胞具有較好的神經分化能力。 結論:無論是人類乳牙牙髓幹細胞或是人類牙根尖幹細胞皆具有分化成神經細胞的能力,但是人類牙根尖幹細胞比人類乳牙牙髓幹細胞具有更好神經分化的能力。我們可以預期利人類牙根尖幹細胞進行神經再生可以得到較佳的結果。 | zh_TW |
dc.description.abstract | Aim:Both Stem cells from human exfoliated deciduous teeth (SHED) and Stem cells from apical papilla (SCAP) are multipotent stem cells. After neural induction, these two kinds of cells could be differentiated into neuron like cells. We hypothesized that SHED and SCAP have the same neurodifferentiation potentials. The purpose of this study is to compare the differences of neurodifferentiation potentials of SHED and SCAP.
Materials and Methods:We cultivated SHED and SCAP in neural induction medium for 0 day, 1 day ,3 days, 7 days , 14 days and 21 days and analyzed cell morphology, cell proliferation, gene expression patterns(RT-PCR) and immunofluorescence before and after differentiation. Results:After 3 days cultivation in neural induction medium, the morphology of SHED and SCAP were changed to neuron like cells. We found that cell proliferation of SHED and SCAP were reduced. After 7 days cultivation in neural induction medium, gene expression patterns (RT-PCR) and Immunofluorescence analysis of the expression Neurofilament demonstrated that both SHED and SCAP were successfully differentiate into neuron cells. However, the Neurofilament gene expression of inducted SCAP was much upregulated then that of inducted SHED. This result indicated that the neurodifferentiation potential of SCAP is higher than that of SHED. Conclusion :Although both SHED and SCAP have potentials for neurodifferentiation but neurodifferentiation potential of SCAP is much better than that of SHED. We can expect that by using SCAP for neurogenesis would get better results. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T18:09:24Z (GMT). No. of bitstreams: 1 ntu-101-R98422025-1.pdf: 3235600 bytes, checksum: 4356ac2e38631d5c42a7c67c6a641e48 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 論文口試委員審定書 i
誌謝 ii 中文摘要 iii ABSTRACT v CONTENTS vii LIST OF FIGURES ix LIST OF TABLES xi Chapter 1 Introduction 1 1.1 Mesenchymal stem cell 1 1.1.1 Stem cells from human exfoliated deciduous teeth (SHED) 2 1.1.2 Stem cells from apical papilla (SCAP): 3 1.2 Neurodifferentiation 5 1.2.1 Markers of neurons 7 Chapter 2 The Purposes and Hypothesis of the Study 9 Chapter 3 Materials and Methods 10 3.1 Isolation and culture of Stem cells from human exfoliated deciduous teeth(SHED) 11 3.2 Isolation and culture of human stem cells from apical papilla (SCAP) 11 3.3 Identification of SHED and SCAP 12 3.4 MTT assay for cell growth analysis 12 3.5 Induction for neurodifferentiation 13 3.6 Identification of neurodifferentiation by Immunofluorescence 13 3.7 RT-PCR analysis 14 3.7.1 Isolation of total RNA 14 3.7.2 RNA Quantitation 14 3.7.3 Reverse Transcription (RT) 15 3.7.4 Polymerase Chain Reaction (PCR) 15 3.8 Statistical analysis 16 Chapter 4 Results 17 4.1 Surface markers of SHED and SCAP 17 4.2 Morphological observation on SHED and SCAP after neural induction 17 4.3 Effects of NDM on cell proliferation of SHED and SCAP-MTT assay 17 4.4 RT-PCR result of SHED and SCAP 18 4.5 RT-PCR Statistical Data of SHED 19 4.6 RT-PCR Statistical Data of SCAP 20 4.7 RT-PCR Statistical Data of SCAP compared with SHED 21 4.8 Immunofluorescence of SHED and SCAP 23 Chapter 5 Discussion 25 5.1 Related factors for neurodifferentiation of SHED and SCAP 25 5.2 Effects of neurodifferentiation on cell morphology change and reducing proliferation 26 5.3 Genes expression during neurodifferentiation of cells 27 5.4 Immunofluorescence of neural induction 29 Chapter 6 Conclusion 30 REFERENCE 31 Figures 36 Table 62 | |
dc.language.iso | en | |
dc.title | 人類乳牙牙髓幹細胞和牙根尖幹細胞神經分化之比較 | zh_TW |
dc.title | Neurodifferentiation potentials of stem cells from human exfoliated deciduous teeth and stem cells from apical papilla | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 郭敏光,李旺祚 | |
dc.subject.keyword | 人類乳牙牙髓幹細胞,人類牙根尖幹細胞,神經分化, | zh_TW |
dc.subject.keyword | Stem cells from human exfoliated deciduous teeth,Stem cells from apical papilla,Neurodifferentiaiton, | en |
dc.relation.page | 78 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2012-07-12 | |
dc.contributor.author-college | 牙醫專業學院 | zh_TW |
dc.contributor.author-dept | 臨床牙醫學研究所 | zh_TW |
顯示於系所單位: | 臨床牙醫學研究所 |
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