Oscillatory Flow Effects on Rat Aortic Smooth Muscle Cells
- Art: Diplomarbeit
- Autor: Joachim Rapp
- Abgabedatum: Februar 1997
- Umfang: 107 Seiten
- Dateigröße: 5,3 MB
- Note: 1,2
- Institution / Hochschule: Clark Atlanta University USA
- ISBN (eBook): 978-3-8324-0048-4
-
ISBN (Paperback) :
978-3-8324-0048-4 P - ISBN (CD) :978-3-8324-0048-4 CD
- Sprache: Englisch
- Prämierung:
- Arbeit zitieren: Rapp, Joachim Februar 1997: Oscillatory Flow Effects on Rat Aortic Smooth Muscle Cells, Hamburg: Diplomica Verlag
- Schlagworte: Arteriosklerose, Glatte Muskelzellen, oszilierende Strömung, Silikon-Substrat, Zellorientierung und -vermehrung
In den Warenkorb
38,00 €
Diplomarbeit von Joachim Rapp
Abstract:
A cell culture System to mimic the circumferential expansion of the arterial wall was supplemented with a flow control System for model enhancement. The given System imposed uniaxial sinusoidal stretch (1 Hz) with a 10 % elongation to an elastic silicone substrate upon which rat aortic smooth muscle cells were cultured. Occurring fluid motion during a stretch experiment caused oscillating shear stress upon the Gell layer of approximately 0.6 dynes/cm2 (60 x 10-3 N/m2) and was controlled by the newly added oscillatory flow System.
Experiments were performed and investigated at 0, 4, and 24 hours. Morphological observations correlated with the results obtained by the initial stretch experiments. A final median angle of orientation of 60° - 70° from the axis of stretch was observed. Both control cultures remained randomly oriented throughout all experiments. Inhibition of cell proliferation alter 4 hours of cyclic stretch, observed by Karen J. Schnetzer could not be confirmed. However, growth related results did correspond to the preceding study in a qualitative manner. Influences of oscillatory flow an SMC growth and morphology were not different to the steady-stretch control. Analysis of results confirmed the assumption made for the earlier culture system, that effects of oscillatory fluid motion occurring during the cyclic stretch experiment could be neglected.
Table of Contents:
| Acknowledgments | V | |
| List of Figures | IX | |
| List of Tables | XI | |
| List of Symbols | XIII | |
| 1. | Introduction | 1 |
| 2. | Background and Literature Review | 3 |
| 2.1 | Arterial Anatomy | 4 |
| 2.2 | Arterial Physiology | 6 |
| 2.3 | Arterial Mechanics | 7 |
| 2.3.1 | Tensile Stress and Arterial Wall Deformation | 7 |
| 2.3.2 | Shear Stress | 8 |
| 2.4 | Arterial Pathology | 10 |
| 2.4.1 | Atherosclerosis | 10 |
| 2.4.2 | Hypertension | 12 |
| 2.4.3 | Smooth Muscle Cells in Arterial Disease | 12 |
| 2.5 | Cell Culture Models of Arterial Mechanics | 13 |
| 2.5.1 | Shear Models | 14 |
| 2.5.2 | Stretch Models | 14 |
| 3. | Materials and Methods | 17 |
| 3.1 | Stretch Chamber | 17 |
| 3.1.1 | Demands on the Stretch System | 18 |
| 3.1.2 | Stretch Chamber Design | 19 |
| 3.2 | Flow Chamber | 20 |
| 3.2.1 | Mathematical Description of the Flow | 22 |
| 3.2.2 | Demands on the Flow Chamber | 27 |
| 3.2.3 | Flow Chamber Design | 27 |
| 3.3 | Elastic Substrate | 31 |
| 3.3.1 | Silicone Membranes | 31 |
| 3.3.2 | Extracellular Matrix | 33 |
| 3.4 | Cell Culture | 35 |
| 3.4.1 | Smooth Muscle Cells | 36 |
| 3.4.2 | Culture. Procedures | 37 |
| 3.5 | Experimental Set-Up | 40 |
| 3.5.1 | Equipment Preparation | 40 |
| 3.5.2 | Substrate Preparation | 41 |
| 3.5.3 | Cell Seeding Method | 43 |
| 3.5.4 | Stretch and Flow Chamber Assembling | 44 |
| 3.5.5 | Experimental Performance | 45 |
| 3.5.6 | Microscopic Visualization and Documentation | 46 |
| 3.5.6.1 | Phase Contrast Microscopy | 46 |
| 3.5.6.2 | Fluorescent Microscopy | 46 |
| 3.5.6.3 | Video Imaging System | 46 |
| 3.6 | Experimental Data Analysis | 48 |
| 3.6.1 | Morphological Analysis | 48 |
| 3.6.2 | Cell Proliferation Analysis | 49 |
| 3.6.2.1 | Proliferating Cell Nuclear Antigen | 50 |
| 3.6.2.2 | Immunofluorescent Staining Techniques | 51 |
| 3.6.2.3 | Total Staining | 52 |
| 4. | Results | 53 |
| 4.1 | Observation Methods and Conditions | 53 |
| 4.2 | Observation of Cell Morphology | 55 |
| 4.2.1 | Cell Shape | 55 |
| 4.2.2 | Cell Orientation | 57 |
| 4.2.3 | Cell Area | 59 |
| 4.3 | Observation of Cell Proliferation | 60 |
| 5. | Discussion | 64 |
| 5.1 | Discussion of the Cell Culture System | 64 |
| 5.1.1 | Stretch Unit | 65 |
| 5.1.2 | Flow Unit | 65 |
| 5.2 | Discussion of the Experimental Results | 66 |
| 5.2.1 | Cell Morphology Data | 66 |
| 5.2.2 | Cell Growth Related Data | 68 |
| 6. | Conclusions | 70 |
| 7 | Appendices | XV |
| 7.1 | Appendix A: Summaries of Data and Statistical Analysis | XV |
| 7.2 | Appendix B: Cell Pictures of Morphological Observations | XXI |
| 7.3 | Appendix C: Detailed Design of the Frame Bars | XXIV |
| 7.4 | Appendix D: Experimental Equipment – Cell Culture Components | XXV |
| 8. | References | XXVII |
In den Warenkorb
38,00 €
Link zur Arbeit:
http://www.diplom.de/ean/9783832400484
Arbeit zitieren:
Rapp, Joachim Februar 1997: Oscillatory Flow Effects on Rat Aortic Smooth Muscle Cells, Hamburg: Diplomica Verlag
Schlagworte:
Arteriosklerose, Glatte Muskelzellen, oszilierende Strömung, Silikon-Substrat, Zellorientierung und -vermehrung
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