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研究生:柯傑
研究生(外文):Chien Ke
論文名稱:膠原蛋白組成調控胎盤衍生幹細胞分化與生長之研究
論文名稱(外文):Study different forms of type I collagen on regulating differentiation and proliferation of human placenta derived multipotent cells
指導教授:陳政男陳政男引用關係張心怡張心怡引用關係
指導教授(外文):Cheng-Nan ChenHsin-I Chang
學位類別:碩士
校院名稱:國立嘉義大學
系所名稱:生化科技學系研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:英文
論文頁數:53
中文關鍵詞:胎盤衍生多功能細胞平滑肌細胞第一型膠原蛋白
外文關鍵詞:Placenta-derived multipotent cellsSmooth muscle cellsType I collagen
相關次數:
  • 被引用被引用:0
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  • 下載下載:117
  • 收藏至我的研究室書目清單書目收藏:1
在動脈壁中,第一型膠原蛋白是構成細胞外基質(ECM)的主要成分,並且可以影響細胞的生長以及分化。本研究的目的是要去探討將胎盤多功能衍生幹細胞(PDMCs)培養在多元膠原蛋白(polymer collagen)和單元膠原蛋白(monomer collagen)上,對於細胞增生和分化有何影響。
胎盤多功能衍生幹細胞培養在多元膠原蛋白以及單元膠原蛋白,並且加入轉型生長因子(transforming growth factor β,TGF-β),平滑肌細胞(SMC)的獨特基因表現型,包括a-smooth muscle actin (a-SMA) 和 calponin會增加。而多元膠原蛋白會增加p21CIP1 和 p27KIP12的基因表現,減少cyclin A, cyclin D1, cyclin-dependent protein kinase 2 (Cdk2)的基因表現;並且使胎盤多功能衍生幹細胞的細胞周期停在 G0/G1期。
因此,多元膠原蛋白具有誘導胎盤多功能衍生幹細胞分化成平滑肌細胞以及在分化成平滑肌細胞的過程中抑制其增生。

Type I collagen constitutes a major portion of the extracellular matrix (ECM) in arterial wall and it is the major substrate for cell growth and differentiation. The goal of this study was to evaluate the differentiation and proliferation of placenta-derived multipotent cells (PDMCs) on polymerized type I collagen fibrils and monomer collagen. PDMCs grown on both polymerized collagen and monomer collagen with transforming growth factor (TGF)- treatment increases the expression of smooth muscle cell (SMC)-specific markers, including a-smooth muscle actin (a-SMA) and calponin. Polymerized collagen increased the expressions of p21CIP1 and p27KIP1; decreased cyclin A, cyclin D1, cyclin-dependent protein kinase 2 (Cdk2); and led to G0/G1 arrest in PDMCs. Thus, polymerized collagen exhibits the great potential in inducing PDMCs differentiation into SMCs, and exerts anti-proliferative effect on PDMC-differentiated SMCs.
Chapter I Introduction …………………………………………… 1
1.1 Introduction of adult stem cell …………………………… 2
1.2 Properties of smooth muscle cell ………………………… 2
1.3 Atherosclerosis and smooth muscle cell ………………… 4
1.4 Molecular mechanisms by differentiation of stem cell …… 4
1.5 Motive …………………………………………………… 5
Chapter II Materials and Methods ……………………………… 7
2.1. Materials ………………………………………………… 8
2.2. Cell culture …………………………………………… 8
2.3. Collagen matrices ……………………………………… 9
2.4. RNA Isolation and Real-time PCR …………………… 9
2.5. Western blot analysis ………………………………… 11
2.6. siRNA Transfection …………………………………… 12
2.7. MTT assay and flow cytometric analysis for cell proliferation ……………………………………………… 12
2.8. Statistical analysis ………………..………………… 13
Chapter III Results ……………………………………………… 14
3.1. Evaluation of PDMC differentiation …………………… 15
3.2. The effect of TGF-β on PDMC differentiation ……… 15
3.3. Proliferation characteristics of PDMC–differentiated SMCs 16
3.4. Comparative analysis of cell cycle regulatory proteins in PDMC–differentiated SMCs …………………….…… 17
3.5. Effect of polymerized collagen on platelet-derived growth factor (PDGF)-BB stimulated PDMC-differentiated SMC proliferation ……………………………………….. 18
3.6. p38 MAPK mediates polymerized collagen-induced differentiation of PDMCs into SMCs …………….…… 19
Chapter IV Discussion ………………………………………… 21
Chapter V Conclusion …………………………………….. 27
References ………………………………………………………… 29
Table 1. Cell cycle analysis of PDMCs cultured on monomer and polymerized collagen. …………………………………… 35
Table 2. Anti-proliferative effect on PDMC-differentiated SMCs by polymerized collagen. …………………………………… 36
Fig. 1. Effect of monomer and polymerized collagen on the expressions of calponin,-SMA and SM-MHC in PDMCs. …………… 37
Fig. 2. Effect of monomer and polymerized collagen on PDMC viability and cell cycle analyses. ………………………………… 39
Fig. 3. Regulation of cell cycle regulatory protein expressions in PDMCs on monomer collagen with TGF- treatment or polymerized collagen. …………………………………………………… 42
Fig. 4. p38 MAPK pathway is required for polymerized collagen-induced calponin and a-SMA expressions. ………………………. 45

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