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研究生：	吳佩蓉 Wu, Pei-Jung
論文名稱：	基質剛性梯度下之NIH3T3纖維母細胞攤附與遷移一維動態模擬 One-dimensional Simulation of Spreading and Migration of NIH3T3 Fibroblast Cells on the Substrate with a Stiffness Gradient
指導教授：	朱銘祥 Ju, Ming-Shuang 林宙晴 Lin, Chou-Ching K.
學位類別：	碩士 Master
系所名稱：	工學院 - 機械工程學系 Department of Mechanical Engineering
論文出版年：	2011
畢業學年度：	100
語文別：	英文
論文頁數：	102
中文關鍵詞：	細胞移動、細胞攤附、數學模型、基質剛性梯度
外文關鍵詞：	cell migration, cell spreading, mathematical model, substrate stiffness
相關次數：	點閱：231 下載：1
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細胞遷移的特性在生理上與傷口癒合、白血球免疫反應、甚至於細胞癌化等都有密切的相關性，而細胞剛性則會影響細胞爬行的運動學特性。因此本研究目的為發展NIH3T3纖維母細胞於基質剛性梯度下之細胞攤附與遷移之一維動態模型，並利用生物力學觀點探討細胞在運動中可能的生理機制。
本研究目的為發展一細胞動態模型，並以系統觀點解釋其生理現象。其中細胞攤附模型整合細胞前伸、集中附著點產生及應力纖維三過程模擬單一細胞從懸浮到攤附的過程。細胞攤附模型中，細胞藉由極化判斷遷移方向，而細胞也會降解後端的集中附著點。基質剛性是影響細胞遷移的重要因素，故本研究提出一製作剛性梯度的方法，使細胞培養於此基質上能產生明顯的運動，並由其運動分析所得到的參數代入遷移模型中。透過模擬能得到細胞的動態行為，並由模擬結果可推測細胞在攤附前期的G-actin濃度會增加以加速攤附；在攤附後期G-actin濃度下降使細胞面積達到飽和。遷移模型部份則呈現細胞在較快運動、較慢運動及往回走的動態情形。由以上生物力學之分析，細胞可不經由生化分析而預測其中之分子性質並模擬出細胞運動的動態情形。

Cell migration plays an important role in the modulation of physiological functions such as wound healing, immunoresponse, and carcinogenesis. A substrate with a stiffness gradient affects the kinematics of cell migration. In this study, a one-dimensional cell spreading and migration model is proposed to investigate the cell motility in the environment of a stiffness gradient by biomechanical analysis.
A one-dimensional cell dynamics model on the substrate with a stiffness gradient is developed in this study, and the physiology is interpreted from system point of view.In the cell spreading model, cell protrusion, focal adhesion formation, and stress fiber formation are integrated to simulate the dynamics from suspension to spreading. In the cell migration model, the movement direction of the modeled cell is determined by polarization, and focal adhesions at the cell rear are degraded. A method for fabricating a substrate with a stiffness gradient is proposed. The parameters obtained from kinematic analysis are substituted into the migration model. Cell dynamics are obtained from simulation, and the G-actin concentration is estimated. The G-actin concentration increases in the early stage of spreading, and then decreases when spreading came to saturate. The simulation results of cell migration shows the dynamics when the cell moved fast, slow, and turned back. The behaviors of cell motility are described by biomechanical analysis, and the molecular properties can be predicted without a biochemical assay.

摘要 I
Abstract II
致謝 III
Table of contents IV
List of figures VIII
List of tables XI
Nomenclature XII
Chapter 1 Introduction 1
1 Background information 1
1.1 Cytoskeleton 1
1.2 Cell migration process 3
1.3 Actin retrograde flow 5
1.4 Categories of stress fibers 6
2 Literature review 7
2.1 Cell biology of cell migration 7
2.2 Mathematical models for cell migration 9
2.3 Effect of substrate stiffness on cell motility 10
3 The statement of purpose 11
4 Structure of the thesis 12
Chapter 2 Materials and Methods	13
1 One-dimensional dynamic models of the NIH3T3 cell 13
1.1 Definitions of dynamics models 13
1.2 Cell spreading model 15
1.3 Cell migration model 19
2 Cell protrusion model 22
2.1 Brownian ratchet model of lamellipodia 22
3 Kinetic model of focal adhesions 25
3.1 Bell’s model 25
3.2 Kong’s microscopic model of focal adhesions 27
3.3 Integrin clustering model	29
4 Models of stress fibers	31
4.1 Two-phase model of lamella 32
4.2 Stress fiber distribution described by Hill equation	34
5 Estimation of G-actin concentration in the cell 35
6 Regulation mechanism of myosin concentration 37
7 Numerical simulation 38
8 Fabrication and measurement of substrate stiffness gradient 39
8.1 Fabrication of substrate with stiffness gradient 39
8.2 Measurement of substrate stiffness 40
9 Cell culture 41
9.1 Medium for NIH3T3 fibroblast cells 41
9.2 Subculture for NIH3T3 fibroblast cells 41
10 Cell spreading and migration experiments	43
10.1 Time-lapse microscopy of cell spreading	43
10.2 Time-lapse microscopy of cell migration	43
10.3 Immunofluorescence 44
10.4 Image process 45
Chapter 3 Results 47
1 Measurement of substrate stiffness	47
2 Experimental results of cell spreading and migration	48
2.1 Experimental results of cell spreading 48
2.2 Results of control group1 (null UV) 52
2.3 Results of control group 2 (full UV) 56
2.4 Results of experimental group (with stiffness gradient) 60
2.5 Immunofluorescence 67
3 Simulation results of focal adhesions kinetic model	70
3.1 Bell’s model	70
3.2 Integrin clustering model	72
4 Simulation results of actin and stress fibers 74
4.1 Brownian ratchet model 74
4.2 Hill equation 76
4.3 Two-phase flow model 77
5 Simulation results of cell spreading model 79
6 Simulation results of cell migration 84
Chapter 4 Discussion 90
1 Migration ability 90
1.1 On control group results 90
1.2 On experimental group results 90
1.3 Comparison of the three group results 91
2 Immunofluorescence results for spreading and migration 91
3 Comparisons with existing models 92
3.1 With Kong’s model	92
3.2 With Dokukina’s migration model 93
4 Physiological interpretation 94
5 Limitations in experiments	95
5.1 Cell division in cell migration experiments 95
5.2 Cell number in migration experiments 97
Chapter 5 Conclusion 100
1 Conclusion 100
2 Future work	100
References  101
                                    

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校內：2014-02-06公開
校外：2014-02-06公開

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